Compounds for the prevention and treatment of diseases and the use thereof

ABSTRACT

Aspects of the invention relate to novel synthetic compounds having binding affinity with galectin proteins.

RELATED APPLICATION(S)

This application is a continuation of U.S. application Ser. No.16/611,609, filed Nov. 7, 2019, which is U.S. national phase applicationunder 35 U.S.C. 371 of PCT International Application No.PCT/US2018/032349, filed on May 11, 2018, which claims the benefit ofand priority to U.S. Provisional Application Ser. No. 62/505,544, filedMay 12, 2017, the entire disclosure of each of which is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

Aspects of the invention relate to compounds, pharmaceuticalcompositions, methods for the manufacturing of compounds and methods fortreatment of various disorders mediated at least in part by one or moregalactose binding proteins also referred to as Galectins.

BACKGROUND OF THE INVENTION

Galectins are a family of S-type lectins that bindbeta-galactose-containing glycoconjugates. To date, fifteen mammaliangalectins have been isolated. Galectins regulate different biologicalprocesses such as cell adhesion, regulation of growth, apoptosis,inflammation, fibrogenesis, tumor development and progression. Galectinshave been shown to be involved in inflammation, fibrosis formation, celladhesion, cell proliferation, metastasis formation, angiogenesis, cancerand immunosuppression.

SUMMARY OF THE INVENTION

Aspects of the invention relate to compounds or compositions comprisinga compound in an acceptable pharmaceutical carrier for parenteral orenteral administration, for use in therapeutic formulations. In someembodiments, the composition can be administered parenterally via anintravenous, subcutaneous, dermal or oral route.

Aspects of the invention relate to compounds or compositions for thetreatment of various disorders in which lectin proteins play a role inthe pathogenesis, including but not limited to, chronic inflammatorydiseases, fibrotic diseases, and cancer. In some embodiments, thecompound is capable of mimicking glycoprotein interactions with lectinsor galectin proteins which are known to modulate the pathophysiologicalpathways leading to inflammation, fibrogenesis, angiogenesis, cancerprogression and metastasis.

According to some aspects of the invention, the compounds comprisepyranosyl and/or furanosyl structures conjugated through an A-M spacerof at least 2 atoms comprising an amide —N(—Ra)—C(═O)—, sulfonamide—N(—H)—S(═O2)-, a methylether —C(—H2)-O— methylester —C(═O)—O—,carbosulfon —C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, Hydrazide —N(—H)—N(—H)— and/or an amino acid.

In some embodiments, the A-M spacer comprises of an amide—N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, a methylether —C(—H2)-O—methylester —C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—, phosphate—O—P(═O)(—OH)—, diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide—C(═O)—NH—NH—, sulfonohydrazide —S(═O)2-NH—NH—, or a phosphonicdihydrazide —P(═O)(—NH—NH2)(NH—NH—) spacer or any combination of theforegoing.

In some embodiments, the spacer is linked to the anomeric carbon of thepyranosyl and/or furanosyl structure.

In some embodiments, the compound comprises organic substituents. Insome embodiments, specific aromatic substitutions can be linked to thegalactose core or the “AM” linker of the anomeric carbon of thepyranosyl and/or furanosyl structures. Such aromatic substitutions canenhance the interaction of the compound with amino acid residues (e.g.Arginine, Tryptophan, Histidine, Glutamic acid etc. . . . ) composingthe carbohydrate-recognition-domains (CRD) of the lectins or with aminoacid residues in the CRD neighborhood and thus strengthen theassociation and binding specificity.

In some embodiments, the organic substituents comprise monosaccharides,disaccharides, oligosaccharides or a heteroglycoside such as iminosugaror thiosugar carbohydrates.

In some embodiments, the compound is a symmetric digalactoside, whereinthe two galactosides are bound by an A-M spacer. Yet in otherembodiments, the compound can be comprised of asymmetric carbohydrates.For example, each of the galactoside can have different aromatic oraliphatic substitutions or heterotatoms derivatives of the galactosewhere the C5 oxygen is replaced with S (5-Thio-D-galactose) or N(5-imino-D-galactose).

Without being bound to the theory, it is believed that A-M spacer renderthe compounds metabolically stable while maintaining the chemical,physical and allosteric characteristics for specific interaction withlectins or galectins known to recognize carbohydrates. In someembodiments, the GalactoAmide and the GalactoSulfonamide of theinvention are metabolically more stable than compounds having anO-glycosidic bond.

Aspects of the invention relate to a compound of formula 1 or apharmaceutically acceptable salt or solvate thereof:

wherein A is selected from the group consisting of NRa, CRb, PRc, andamino acid,

wherein M is selected from the group consisting of NRa, CRb, PRc, ORd,SRe amino acid, and hydrophobic hydrocarbons derivatives includingheterocyclic substitutions of 3 or more atoms,

wherein Ra is selected from the group consisting of H, H2, CH3, COOH,NH2, COMe, halogen and combinations thereof,

wherein Rb is selected from the group consisting of H, H2, O, OH, CH3,COOH, NH2, COMe, halogen and combinations thereof,

wherein Rc is selected from the group consisting of O2, PO2, OH, halogenand combinations thereof,

wherein Rd is selected from the group consisting of H and CH3, andcombinations thereof,

wherein Re is selected from the group consisting of OH, O2, S, halogenand combinations thereof,

wherein B is OH, NH2, NHAc, or NH-alkyl wherein the alkyl comprises 1 to18 Carbons,

wherein W is selected from the group consisting of O, S, CH2, NH, or Se,

wherein Y is selected from the group consisting of O, S, NH, CH2, Se, S,P, amino acid, and hydrophobic linear and cyclic hydrophobichydrocarbons derivatives including heterocyclic substitutions ofmolecular weight of about 50-200 D and combinations thereof,

wherein R₁, R₂, and R₃ are independently selected from the groupconsisting of H, O2, CO, NH2, SO2, SO, PO2, PO, CH3, linear hydrocarbon,and cyclic hydrocarbon, and

wherein the hydrocarbon is one of a) an alkyl group of at least 3carbons, an alkenyl group of at least 3 carbons, an alkyl group of atleast 3 carbons substituted with a carboxy group, an alkenyl group of atleast 3 carbons substituted with a carboxy group, an alkyl group of atleast 3 carbons substituted with an amino group, an alkenyl group of atleast 3 carbons substituted with an amino group, an alkyl group of atleast 3 carbons substituted with both an amino and a carboxy group, analkenyl group of at least 3 carbons substituted with both an amino and acarboxy group, and an alkyl group substituted with one or more halogens,b) a phenyl group substituted with at least one carboxy group, a phenylgroup substituted with at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group, c) a naphthyl group, a naphthylgroup substituted with at least one carboxy group, a naphthyl groupsubstituted with at least one halogen, a naphthyl group substituted withat least one alkoxy group, a naphthyl group substituted with at leastone nitro group, a naphthyl group substituted with at least one sulfogroup, a naphthyl group substituted With at least one amino group, anaphthyl group substituted with at least one alkylamino group, anaphthyl group substituted with at least one dialkylamino group, anaphthyl group substituted with at least one hydroxy group, a naphthylgroup substituted with at least one carbonyl group and a naphthyl groupsubstituted with at least one substituted carbonyl group, d) aheteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group, and e) asaccharide, a substituted saccharide, D-galactose, Deoxygalactose,substituted D-Galctose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose,hydrogen, an alkyl group, an alkenyl group, an aryl group, a heteroarylgroup, and a heterocycle and derivatives, an amino group, a substitutedamino group, an imino group, or a substituted imino group.

In some embodiments, wherein A-M is representing a spacer of at least 2atoms comprising an amide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, amethylether —C(—H2)-O— methylester —C(═O)—O—, carbosulfon—C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, Hydrazide —N(—H)—N(—H)—, and amino acid, orcombinations thereof.

In some embodiments, the A-M spacer comprises of an amide—N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, a methylether —C(—H2)-O—methylester —C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—, phosphate—O—P(═O)(—OH)—, diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide—C(═O)—NH—NH—, sulfonohydrazide —S(═O)2-NH—NH—, or a phosphonicdihydrazide —P(═O)(—NH—NH2)(NH—NH—) spacer or any combination of theforegoing.

In some embodiments, the A-M spacer 2 or more atoms linked by single ordouble bond: C—C, C═C, C—P, C—N, C—O, N—C, N—N, N═N, N—S, N—P, S—N, P—O,O—P, S—C, S—N, S—S or combination thereof.

In some embodiments, the A-M spacer comprises PO2 or PO2-PO2 bond linkedto the anomeric carbon and to one or more atoms such as C or N or O orS. In some embodiments, C or N is linked to the anomeric carbon and PO2or PO2-PO2 is linked to C or N.

In some embodiments, the A-M is methylamide linked R1, R2 isN′-methylamide-3,4-difluorobenzene and Y—R₁ is triazole-3-fluorobenzene

In some embodiments, the A-M spacer is linked to a galactose, a hydroxylcyclohexane, an aromatic moiety, an alkyl group, an aryl group, an aminegroup, or amide group.

In some embodiments, the A-M spacer symmetrically links two galactosidesor substituted derivatives thereof.

In some embodiments, the A-M spacer asymmetrically links twogalactosides or substituted derivatives thereof.

In some embodiments, the anomeric carbon of the galactoside has a spacerof 2 or more atoms linked by single or double bond: C—C, C═C, C—P, C—N,C—O, N—C, N—N, N═N, N—S, N—P, S—N, P—O, O—P, or combination thereof.

Aspects of the invention relate to a compound or a pharmaceuticallyacceptable salt or solvate thereof having Formula 2:

wherein A is independently selected the group consisting of NRa, CRb,PRc, or amino acid,

wherein M is independently selected from the group consisting of NRa,CRb, PRc, ORd, SRe amino acid, or hydrophobic hydrocarbons derivativesincluding heterocyclic substitutions of 3 or more atoms,

wherein Ra is selected from the group consisting of H, H2, CH3, COOH,NH2, COMe, halogen and combinations thereof,

wherein Rb is selected from the group consisting of H, H2, O, OH, CH3,COOH, NH2, COMe, halogen and combinations thereof,

wherein Rc is selected from the group consisting of O2, PO2, OH, halogenand combinations thereof,

wherein Rd is selected from the group consisting of H and CH3,

wherein Re is selected from the group consisting of OH, O2, S, halogenand combinations thereof,

wherein B is OH, NH2, NHAc, or NH-alkyl wherein the alkyl comprises 1 to18 Carbons,

wherein W is selected from the group consisting of O, S, CH2, NH, andSe,

wherein X is selected from the group consisting of O, N, S, CH2, NH, andPO2,

wherein Y and Z are selected from the group consisting of O, S, C, NH,CH2, Se, S, P, amino acid, and hydrophobic linear and cyclic hydrophobichydrocarbons derivatives including heterocyclic substitutions ofmolecular weight of about 50-200 D and combinations thereof,

wherein R1, R2, R3, are independently selected from the group consistingof CO, O2, SO2, SO, PO2, PO, CH, Hydrogen, hydrophobic linearhydrocarbon, and hydrophobic cyclic hydrocarbon, wherein the hydrocarbonis one of:

a) an alkyl group of at least 3 carbons, an alkenyl group of at least 3carbons, an alkyl group of at least 3 carbons substituted with a carboxygroup, an alkenyl group of at least 3 carbons substituted with a carboxygroup, an alkyl group of at least 3 carbons substituted with an aminogroup, an alkenyl group of at least 3 carbons substituted With an aminogroup, an alkyl group of at least 3 carbons substituted with both anamino and a carboxy group, an alkenyl group of at least 3 carbonssubstituted with both an amino and a carboxy group, and an alkyl groupsubstituted with one or more halogens;b) a phenyl group substituted with at least one car boxy group, a phenylgroup substituted With at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group;c) a naphthyl group, a naphthyl group substituted with at least onecarboxy group, a naphthyl group substituted with at least one halogen, anaphthyl group substituted with at least one alkoxy group, a naphthylgroup substituted with at least one nitro group, a naphthyl groupsubstituted with at least one sulfo group, a naphthyl group substitutedWith at least one amino group, a naphthyl group substituted with atleast one alkylamino group, a naphthyl group substituted with at leastone dialkylamino group, a naphthyl group substituted with at least onehydroxy group, a naphthyl group substituted with at least one carbonylgroup and a naphthyl group substituted with at least one substitutedcarbonyl group; andd) a heteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group;e) saccharide, a substituted saccharide, D-galactose, substitutedD-galactose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose, hydrogen,an alkyl group, an alkenyl group, an aryl group, a heteroaryl group, anda heterocycle and derivatives, an amino group, a substituted aminogroup, an imino group, or a substituted imino group.

In some embodiments, A-M represents a spacer of at least 2 atomscomprising an amide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, amethylether —C(—H2)—O— methylester —C(═O)—O—, carbosulfon—C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide —C(═O)—NH—NH—,sulfonohydrazide —S(═O)2-NH—NH—, and phosphonic dihydrazide—P(═O)(—NH—NH2)(NH—NH—) or combinations thereof.

In some embodiments, the A-M spacer 2 or more atoms linked by single ordouble bond: C—C, C═C, C—P, C—N, C—O, N—C, N—N, N═N, N—S, N—P, S—N, P—O,O—P, S—C, S—N, S—S or combination thereof.

In some embodiments, the A-M spacer of at least 2 atoms has a rotationalfreedom and length configured to allow an interaction of about 1 nM toabout 50 μM to a galectin CRD epitope.

In some embodiments, the hydrophobic linear and cyclic hydrocarbonsincluding heterocyclic substitutions have a molecule weight of about 50to 200 D.

Aspects of the invention relate to a compound or a pharmaceuticallyacceptable salt or solvate thereof having formula of Table 1.

In some embodiments, the compound is in a free form. In someembodiments, the free form is an anhydrate. In some embodiments, thefree form is a solvate, such as a hydrate.

In some embodiments, the compound of Formula (1) or Formula (2) is acrystalline form.

Some aspects of the invention relate to a compound of formula (1) foruse as a therapeutic agent in a mammal, such as a human.

Some aspects of the invention relate to a pharmaceutical compositioncomprising the compound of Formula (1) or Formula (2) and optionally apharmaceutically acceptable additive, such as carrier or excipient.

In some embodiments, the compounds of the invention bind to one or moregalectins. In some embodiments, the compound binds to Galectin-3,Galectin-1, Galectin 8, and/or Galectin 9.

In some embodiments, the compounds of the invention have highselectivity and affinity for Galectin-3. In some embodiments, thecompounds of the invention have an affinity of about 1 nM to about 50 μMfor Galectin-3.

Aspects of the invention relate to compositions comprising the compoundof the invention. In some embodiments, the composition comprises atherapeutically effective amount of the compound and a pharmaceuticallyacceptable adjuvant, excipient, formulation carrier or combinationsthereof. In some embodiments, the composition comprises atherapeutically effective amount of the compound and of ananti-inflammatory drug, vitamin, pharmaceutical drug, nutraceuticaldrug, supplement, or combinations thereof.

Aspects of the invention relate to compounds, composition and methodsthat can be used in the treatment of diseases in a subject in needthereof. Aspects of the invention relate to compounds, composition andmethods that can be used in the treatment of diseases in which galectinsare at least in part involved in the pathogenesis.

Aspects of the invention relate to methods of treatment of a disease ina subject in need thereof. In some embodiments, the subject is a mammal.In some embodiments, the subject is a human.

In some embodiments, the compounds, composition and methods can be usedin the treatment of nonalcoholic steatohepatitis with or without liverfibrosis, inflammatory and autoimmune disorders, neoplastic conditionsor cancers.

In some embodiments, the compounds, composition and methods can be usedin the treatment of liver fibrosis, kidney fibrosis, lung fibrosis, orheart fibrosis.

In some embodiments, the composition or the compound is capable ofenhancing anti-fibrosis activity in organs, including but not limitedto, liver, kidney, lung, and heart.

In some embodiments, the compounds, composition and methods can be usedin treatment of inflammatory disorders of the vasculature includingatherosclerosis and pulmonary hypertension.

In some embodiments, the compounds, composition and methods can be usedin the treatment of heart disorders including heart failure,arrhythmias, and uremic cardiomyopathy.

In some embodiments, the compounds, composition and methods can be usedin the treatment of kidney diseases including glomerulopathies andinterstitial nephritis.

In some embodiments, the compounds, composition and methods can be usedin the treatment of inflammatory, proliferative and fibrotic skindisorders, including but not limited to, psoriasis and scleroderma.

In some embodiments, the invention relates to a method of treatinginflammatory and fibrotic disorders in which galectins are at least inpart involved in the pathogenesis, by enhancing anti-fibrosis activityin organs, including, but not limited to, liver, kidney, lung, andheart.

In some embodiments, the invention relates to a composition or acompound that has a therapeutic activity to treat nonalcoholicsteatohepatitis (NASH). In other aspects, the invention relates to amethod to reduce the pathology and disease activity associated withnonalcoholic steatohepatitis (NASH).

In some embodiments, the invention relates to compounds, composition andmethods for treating inflammatory and autoimmune disorders in whichgalectins are at least in part involved in the pathogenesis includingbut not limited to arthritis, rheumatoid arthritis, asthma, andinflammatory bowel disease.

In some embodiments, the invention relates to a composition or acompound to treat neoplastic conditions (e.g. benign or malignantneoplastic diseases) in which galectins are at least in part involved inthe pathogenesis by inhibiting processes promoted by the increase ingalectins. In some embodiments, the invention relates a method oftreating neoplastic conditions (e.g. benign or malignant neoplasticdiseases) in which galectins are at least in part involved in thepathogenesis by inhibiting processes promoted by the increase ingalectins. In some embodiments, the composition or the compound can beused to treat or prevent tumor cell invasion, metastasis, andneovascularization. In some embodiments, the composition or the compoundcan be used to treat primary and secondary cancers.

In some embodiments, a therapeutically effective amount of the compoundor of the composition can be compatible and effective in combinationwith a therapeutically effective amount of anti-inflammatory drugs,vitamins, other pharmaceuticals and nutraceuticals drugs or supplement,or combinations thereof without limitation.

Some aspects of the invention relate to a compound of Formula (1)Formula or (2) for use in a method for treating a disorder relating tothe binding of a galectin. Some aspects of the invention relate to acompound of Formula (1) or Formula (2) for use in a method for treatinga disorder relating to the binding of galectin-3 to a ligand.

Some aspects of the invention relate to a method for treatment of adisorder relating to the binding of a galectin, such as galectin-3, to aligand in a human, wherein the method comprises administering atherapeutically effective amount of at least one compound of Formula (1)or Formula (2) to a human in need thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

The present invention will be further explained with reference to theattached drawings, wherein like structures are referred to by likenumerals throughout the several views. The drawings shown are notnecessarily to scale, with emphasis instead generally being placed uponillustrating the principles of the present invention.

FIG. 1A depicts a high-definition 3D structure of galectin-3Carbohydrate Recognition Domain (CRD) binding pocket with 3 potentialsites of interaction.

FIG. 1B depicts the CRD pocket location in the Galectin-3 C-terminalwith bound lactose unit.

FIG. 2 depicts a map of the Galectin-3 CRD site vicinity—potentialcooperative amino-acids for enhanced binding.

FIG. 3A depicts in-silico 3D model predicted docking pose of aGalactoamideSuccinimide linked Compound according to some embodiments.

FIG. 3B depicts in-silico 3D model predicted docking pose of aGalactoAmide linked compound according to some embodiments.

FIGS. 4A-4K depict the synthesis of exemplary GalactoAmide compoundsaccording to some embodiments.

FIG. 5A depicts the inhibition of galectin binding moiety using aspecific anti-Galectin-3 monoclonal antibodies binding assay (ELISAformat) according to some embodiments.

FIG. 5B depicts the inhibition of galectin using an integrin-galectin-3functional assay (CRD ELISA format) according to some embodiments.

FIG. 6A depicts a Fluorescence Resonance Energy Transfer analyticalassay (FRET Format) for screening anti-galectin active compoundsaccording to some embodiments.

FIG. 6B depicts a Fluorescent Polarization Assay Format which detectscompounds that bind specifically to the CRD according to someembodiments.

FIG. 7A depicts correlation between the ELISA MAb and the ELISA Integrinassays for multiple GalactoAmides derivatives.

FIG. 7B provides examples of Compounds IC50 (Inhibition concentration50% score) by the ELISA Integrin-Gal-3 and ELISA MAb-Gal-3 assaysaccording to some embodiments.

FIG. 8A shows reduction of Fluorescent Polarization of a CRD specificbinding of fluorescent ligand by compounds (600 series) according tosome embodiments.

FIG. 8B shows examples of target inhibition of Galectin-3 interactionwith Integrin-aMB2 by compounds according to some embodiments of theinvention (600 series).

FIG. 9 shows examples of target inhibition according to some embodimentsby compounds of the invention (600 series) of Galectin-3 interactionwith Integrin-aVB6.

FIGS. 10A and 10B show examples of Inhibition of the cytokine MCP-1secretion by Inflammatory Macrophages (LPS stressed THP-1 cell culture)by compounds and compound IC50 (600 series) according to someembodiments of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Detailed embodiments of the present invention are disclosed herein;however, it is to be understood that the disclosed embodiments aremerely illustrative of the invention that may be embodied in variousforms. In addition, each of the examples given in connection with thevarious embodiments of the invention is intended to be illustrative, andnot restrictive. Further, the figures are not necessarily to scale, somefeatures may be exaggerated to show details of particular components. Inaddition, any measurements, specifications and the like shown in thefigures are intended to be illustrative, and not restrictive. Therefore,specific structural and functional details disclosed herein are not tobe interpreted as limiting, but merely as a representative basis forteaching one skilled in the art to variously employ the presentinvention.

Citation of documents herein is not intended as an admission that any ofthe documents cited herein is pertinent prior art, or an admission thatthe cited documents are considered material to the patentability of theclaims of the present application.

Throughout the specification and claims, the following terms take themeanings explicitly associated herein, unless the context clearlydictates otherwise. The phrases “in one embodiment” and “in someembodiments” as used herein do not necessarily refer to the sameembodiment(s), though it may. Furthermore, the phrases “in anotherembodiment” and “in some other embodiments” as used herein do notnecessarily refer to a different embodiment, although it may. Thus, asdescribed below, various embodiments of the invention may be readilycombined, without departing from the scope or spirit of the invention.

In addition, as used herein, the term “or” is an inclusive “or”operator, and is equivalent to the term “and/or,” unless the contextclearly dictates otherwise. The term “based on” is not exclusive andallows for additional factors not described, unless the context clearlydictates otherwise. In addition, throughout the specification, themeaning of “a,” “an,” and “the” include plural references.

Unless otherwise specified, all percentages expressed herein areweight/weight.

Aspects of the invention relate to compositions of mono, disaccharidesand oligosaccharides of Galactose (or heteroglycoside) core bound to an“amide” or “sulfonamide” linkage on the anomeric carbon of the Galactose(or heteroglycoside). In some embodiments, the “AM” containing moleculesrender them metabolically active while maintaining the chemical,physical and allosteric characteristics for specific interaction withlectins known to recognize carbohydrates. In some embodiments, thespecific aromatic substitutions added to the galactose core furtherenhance the affinity of the “amide” bound pyranosyl and/or furanosylstructures by enhancing their interaction with amino acid residues (e.g.Arginine, Tryptophan, Histidine, Glutamic acid etc. . . . ) composingthe carbohydrate-recognition-domains (CRD) of the lectins and thusstrengthen the association and binding specificity.

Galectins

Galectins (also known as galaptins or S-lectins) are a family of lectinswhich bind beta-galactoside. Galectin as a general name was proposed in1994 for a family of animal lectins (Barondes, S. H., et al.: Galectins:a family of animal beta-galactoside-binding lectins. Cell 76, 597-598,1994), The family is defined by having at least one characteristiccarbohydrate recognition domain (CRD) with an affinity forbeta-galactosides and sharing certain sequence elements. Furtherstructural characterization segments the galectins into three subgroupsincluding: (1) galectins having a single CRD, (2) galectins having twoCRDs joined by a linker peptide, and (3) a group with one member(galectin-3) which has one CRD joined to a different type of N-terminaldomain. The galectin carbohydrate recognition domain is a beta-sandwichof about 135 amino acids. The two sheets are slightly bent with 6strands forming the concave side, also called the S-face, and 5 strandsforming the convex side, the F-face). The concave side forms a groove inwhich carbohydrate is bound (Leffler H, Carlsson S, Hedlund M, Qian Y,Poirier F (2004). “Introduction to galectins”. Glycoconj. J. 19 (7-9):433-40).

A wide variety of biological phenomena have been shown to be related togalectins, including development, differentiation, morphogenesis, tumormetastasis, apoptosis, RNA splicing, and many others.

Generally, the carbohydrate domain binds to galactose residuesassociated with glycoproteins. Galectins show an affinity for galactoseresidues attached to other organic compounds, such as in lactose[(β-D-Galactosido)-D-glucose], N-acetyl-lactosamine,poly-N-acetyllactosamine, galactomannans, or fragments of pectins.However, it should be noted that galactose by itself does not bind togalectins.

Plant polysaccharides like pectin and modified pectin have been shown tobind to galectin proteins presumably on the basis of containinggalactose residues that are presented in the context of a macromolecule,in this case a complex carbohydrate rather than a glycoprotein in thecase of animal cells.

At least fifteen mammalian galectin proteins have been identified whichhave one or two carbohydrate domain in tandem.

Galectin proteins are found in the intracellular space where they havebeen assigned a number of functions and they are also are secreted intothe extracellular space where they have different functions. In theextracellular space, galectin proteins can have multiple functions thatare mediated by their interaction with galactose containingglycoproteins including promoting interactions between glycoproteinsthat may modulate function or, in the case of integral membraneglycoprotein receptors, modification of cellular signaling (Sato et al“Galectins as danger signals in host-pathogen and host-tumorinteractions: new members of the growing group of “Alarmins.” In“Galectins,” (Klyosov, et al eds.), John Wiley and Sons, 115-145, 2008,Liu et al “Galectins in acute and chronic inflammation,” Ann. N.Y. Acad.Sci. 1253: 80-91, 2012). Galectin proteins in the extracellular spacecan additionally promote cell-cell and cell matrix interactions (Wang etal., “Nuclear and cytoplasmic localization of galectin-1 and galectin-3and their roles in pre-mRNA splicing.” In “Galectins” (Klyosov et aleds.), John Wiley and Sons, 87-95, 2008). In regards to intracellularspace, galectin functions appear to be more related to protein-proteininteractions, although intracellular vesicle trafficking appears to berelated to interaction with glycoproteins.

Galectins have been shown to have domains which promotehomodimerization. Thus, galectins are capable of acting as a “molecularglue” between glycoproteins. Galectins are found in multiple cellularcompartments, including the nucleus and cytoplasm, and are secreted intothe extracellular space where they interact with cell surface andextracellular matrix glycoproteins. The mechanism of molecularinteractions can depend on the localization. While galectins caninteract with glycoproteins in the extracellular space, the interactionsof galectin with other proteins in the intracellular space generallyoccurs via protein domains. In the extracellular space the associationof cell surface receptors may increase or decrease receptor signaling orthe ability to interact with ligands.

Galectin proteins are markedly increased in a number of animal and humandisease states, including but not limited to diseases associated withinflammation, fibrosis, autoimmunity, and neoplasia. Galectins have beendirectly implicated in the disease pathogenesis, as described below. Forexample, diseases states that may be dependent on galectins include, butare not limited to, acute and chronic inflammation, allergic disorders,asthma, dermatitis, autoimmune disease, inflammatory and degenerativearthritis, immune-mediated neurological disease, fibrosis of multipleorgans (including but not limited to liver, lung, kidney, pancreas, andheart), inflammatory bowel disease, atherosclerosis, heart failure,ocular inflammatory disease, a large variety of cancers.

In addition to disease states, galectins are important regulatorymolecules in modulating the response of immune cells to vaccination,exogenous pathogens and cancer cells.

One of skill in the art will appreciate that compounds that can bind togalectins and/or alter galectin's affinity for glycoproteins, reducehetero- or homo-typic interactions between galectins, or otherwise alterthe function, synthesis, or metabolism of galectin proteins may haveimportant therapeutic effects in galectin-dependent diseases.

Galectin proteins, such as galectin-1 and galectin-3 have been shown tobe markedly increased in inflammation, fibrotic disorders, and neoplasia(Ito et al. “Galectin-1 as a potent target for cancer therapy: role inthe tumor microenvironment”, Cancer Metastasis Rev. PMID: 22706847(2012), Nangia-Makker et al. Galectin-3 binding and metastasis,” MethodsMol. Biol. 878: 251-266, 2012, Canesin et al. Galectin-3 expression isassociated with bladder cancer progression and clinical outcome,” TumourBiol. 31: 277-285, 2010, Wanninger et al. “Systemic and hepatic veingalectin-3 are increased in patients with alcoholic liver cirrhosis andnegatively correlate with liver function,” Cytokine. 55: 435-40, 2011).Moreover, experiments have shown that galectins, particularly galectin-1(gal-1) and galectin-3 (gal-3), are directly involved in thepathogenesis of these classes of disease (Toussaint et al., “Galectin-1,a gene preferentially expressed at the tumor margin, promotesglioblastoma cell invasion.”, Mol. Cancer. 11:32, 2012, Liu et al 2012,Newlaczyl et al., “Galectin-3—a jack-of-all-trades in cancer,” CancerLett. 313: 123-128, 2011, Banh et al., “Tumor galectin-1 mediates tumorgrowth and metastasis through regulation of T-cell apoptosis,” CancerRes. 71: 4423-31, 2011, Lefranc et al., “Galectin-1 mediated biochemicalcontrols of melanoma and glioma aggressive behavior,” World J. Biol.Chem. 2: 193-201, 2011, Forsman et al., “Galectin 3 aggravates jointinflammation and destruction in antigen-induced arthritis,” ArthritisReum. 63: 445-454, 2011, de Boer et al., “Galectin-3 in cardiacremodeling and heart failure,” Curr. Heart Fail. Rep. 7, 1-8, 2010,Ueland et al., “Galectin-3 in heart failure: high levels are associatedwith all-cause mortality,” Int J. Cardiol. 150: 361-364, 2011, Ohshimaet al., “Galectin 3 and its binding protein in rheumatoid arthritis,”Arthritis Rheum. 48: 2788-2795, 2003).

High levels of serum Galectin-3 have been shown to be associated withsome human diseases, such as a more aggressive form of heart failure,which make identification of high-risk patients using galectin-3 testingan important part of patient care. Galectin-3 testing may be useful inhelping physicians determine which patients are at higher risk ofhospitalization or death. For example, the BGM Galectin-3@ Test is an invitro diagnostic device that quantitatively measures galectin-3 in serumor plasma and can be used in conjunction with clinical evaluation as anaid in assessing the prognosis of patients diagnosed with chronic heartfailure. Measure of the concentration of endogenous protein galectin-3can be used to predict or monitor disease progression or therapeuticefficacy in patients treated with cardiac resynchronization therapy (seeU.S. Pat. No. 8,672,857, which is incorporated herein by reference inits entirety).

Galectin-8 (gal-8) has been shown to be over-expressed in lungcarcinomas and is in the invasive regions of xenografted glioblastomas.

Galectin-9 (gal-9) is believed to be involved in the control of lesionsarising from immunoinflammatory diseases, and be generally implicated ininflammation. Gal-9 appears to mediate apoptosis in certain activatedcells.

Aspects of the invention relate to compounds that bind galectinsinvolved in human disorders, such as inflammatory diseases, fibroticdiseases, neoplastic diseases or combinations thereof. In someembodiments, the compounds bind galectins, such as galectin-1 (gal-1),galectin-3 (gal-3), galectin-8 (gal-8) and/or galectin-9 (gal-9).

Galectin Inhibitors

Natural oligosaccharide ligands capable of binding to galectin-1 and/orgalectin-3, for example, modified forms of pectins and galactomannanderived from Guar-gum have been described (see WO 2013040316, US20110294755, WO 2015138438). Synthetic digalactosides like lactose,N-acetyllactosamine (LacNAc) and thiolactose effective against pulmonaryfibrosis and other fibrotic disease (WO 2014067986 A1).

Advances in protein crystallography and availability of high definition3D structure of the carbohydrate recognition domain (CRD) of manygalectins have generated many derivatives with enhanced affinity to theCRD having a greater affinity than galactose or lactose (WO 2014067986A). These compounds were shown to be effective for treatment of ananimal model of lung fibrosis which is thought to mimic human idiopathicpulmonary fibrosis (IPF). For example a thio-digalactopyranosylsubstituted with 3-fiuorophenyl-2,3-triazol groups (TD-139) has beenreported to bind to galectin 3 and to be effective in in a mouse modelof lung fibrosis. The compound required pulmonary administration usingintra-tracheal instillation or nebulizers (see U.S. Pat. Nos. 8,703,720,7,700,763, 7,638,623 U.S. Pat. No. 7,230,096).

Aspects of the invention relates to novel compounds that mimic thenatural ligand of galectin proteins. In some embodiments, the compoundmimics the natural ligand of galectin-3. In some embodiments, thecompound mimics the natural ligand of galectin-1.

In some embodiments, the compound has a mono, di or oligomer structurecomposed of Galactose-AM core bound to the anomeric carbon on thegalactose and which serves as a linker to the rest of the molecule. Insome embodiments, the Galactose-AM core may be bound to othersaccharide/amino acid/acids/group that bind galectin CRD (as shown inFIG. 1A, 1B) in the high definition 3D structure of galectin-3) andtogether can enhance the compound's affinity to the CRD. In someembodiments, the Galactose-AM core may be bound to othersaccharide/amino acid/acid/group that bind in “site B” of the galectinCRD (as shown in FIGS. 1A, 1B & FIG. 2 in the high definition 3Dstructure of galectin-3) and together can enhance the compound'saffinity to the CRD.

According to some aspects, the compounds can have substitutions thatinteract with site A and/or site C to further improve the associationwith the CRD and enhance their potential as a therapeutic targeted togalectin-dependent pathology. In some embodiments, the substituents canbe selected through in-silico analysis (computer assisted molecularmodeling) as described herein. In some embodiments, the substituents canbe further screened using binding assay with the galectin protein ofinterest. For example, the compounds can be screened using a galectin-3binding assay and/or an in-vitro inflammatory and fibrotic model ofactivated cultured macrophages (see Macrophage polarization minireview,AbD Serotec).

According to some aspects, the compounds comprise one or more specificsubstitutions of the core Galactose-AM. For example, the coreGalactose-AM can be substituted with specific substituents that interactwith residues located within the CRD. Such substituents can dramaticallyincrease the association and potential potency of the compound as wellas the ‘drugability’ characteristic (FIG. 3A, 3B)

Galactoside Compounds

Most “amide” and “sulfon” compounds, organic and inorganic, are readilyabsorbed from the diet and transported to the liver—the prime organ formetabolism. The general metabolism of “amide” compounds follows threemajor routes depending on the chemical properties, that is, redox-active“amide” compounds, precursors of methylamide and conjugation with aminoacids.

AM Spacer

Aspects of the invention relates to compounds comprising pyranosyland/or furanosyl galactose structures bound to an “A-M” spacer on theanomeric carbon of the pyranosyl and/or furanosyl.

In some embodiments, wherein A-M is representing a spacer of at least 2atoms comprising an amide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, amethylether —C(—H2)-O— methylester —C(═O)—O—, carbosulfon—C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, Hydrazide —N(—H)—N(—H)—, and amino acid, orcombinations thereof.

In some embodiments, the A-M spacer comprises an amide —N(—Ra)—C(═O)—,sulfonamide —N(—H)—S(═O2)-, a methylether —C(—H2)-O— methylester—C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—,diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide —C(═O)—NH—NH—,sulfonohydrazide —S(═O)2-NH—NH—, or a phosphonic dihydrazide—P(═O)(—NH—NH2)(NH—NH—) spacer or any combination of the foregoing.

In some embodiments, the A-M spacer comprises 2 or more atoms linked bysingle or double bond: C—C, C═C, C—P, C—N, C—O, N—C, N—N, N═N, N—S, N—P,S—N, P—O, O—P, S—C, S—N, S—S or combination thereof.

In some embodiments, the A-M spacer comprises PO2 or PO2-PO2 bond linkedto the anomeric carbon and to one or more atoms such as C or N or O orS. In some embodiments, C or N is linked to the anomeric carbon and PO2or PO2-PO2 is linked to C or N.

Without being bound to the theory, A-M is representing a spacer of atleast 2 atoms that has more rotational freedom and length thus affordingcloser and tighter interaction to the galectin CRD epitope andsurrounding aminoacids sites. Spacers like an amide —N(—Ra)—C(═O)—,sulfonamide —N(—H)—S(═O2)-, a methylether —C(—H2)-O— methylester—C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—,diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide —C(═O)—NH—NH—,sulfonohydrazide —S(═O)2-NH—NH—, and phosphonic dihydrazide—P(═O)(—NH—NH2)(NH—NH—) augment the interaction with the galectin.

Aspects of the invention relates to compounds comprising pyranosyland/or furanosyl galactose structures bound to an “amide” or “sulfon”type structures on the anomeric carbon of the pyranosyl and/orfuranosyl. As used herein an amide bond referes to a C—N bond(R—C(O)—NH—R). In some embodiments, the amide bond can be a sulfonamidebond. In some embodiments, the sulfon bond can have the genera formulaR—S(═O)₂—R′. As used herein an amide bond refers to a C—N bond(R—C(O)—NH—R). In some embodiments, the amide bond can be a N—SO2(sulfonamide bond). or the genera R—N—S(═O)₂—R′. In some embodiments,the C—SO2 (sulfon bond) can have the genera formula R—C—S(═O)₂—R′.

In some embodiments, specific aromatic substitutions can be added to thegalactose core or heteroglycoside core to further enhance the affinityof the “amide” bound pyranosyl and/or furanosyl structures. Sucharomatic substitutions can enhance the interaction of the compound withamino acid residues (e.g. Arginine, Tryptophan, Histidine, Glutamic acidetc. . . . ) composing the carbohydrate-recognition-domains (CRD) of thelectins and thus strengthen the association and binding specificity.

In some embodiments, the compound comprises monosaccharides,disaccharides and oligosaccharides of galactose or a heteroglycosidecore bound to an “amide” or “sulfon” atom on the anomeric carbon of thegalactose or of the heteroglycoside.

In some embodiments, the compound is a symmetric digalactoside whereinthe two galactosides are bound by one or more “amide” and/or “sulfon”bonds. In some embodiments, the compound is a symmetric digalactosidewherein the two galactosides are bound by one or more sulfonamide bonds.In some embodiments, the compound is a symmetric digalactoside whereinthe two galactosides are bound by one or more “amide” bonds and whereinthe “amide” is bound to the anomeric carbon of the galactose. In someembodiments, the compound is a symmetric digalactoside wherein the twogalactosides are bound by one or more “amide” bonds and one or moresulfon bonds and wherein the “amide” is bound to the anomeric carbon ofthe galactose. Yet in other embodiments, the compound can be anasymmetric digalactoside. For example, the compound can have differentaromatic or aliphatic substitutions on the galactose core.

In some embodiments, the compound is asymmetric galactoside wherein asingle galactoside having one or more “amide” or “sulfon” on theanomeric carbon of the galactose. In some embodiments, the galactosidehas one or more “amide” bound to the anomeric carbon of the galactoseand one or more sulfur bound to the “amide”. In some embodiments, thecompound can have different aromatic or aliphatic substitutions on thegalactose core.

Without being bound to the theory, it is believed that the compoundscontaining the AM linkage render the compound metabolically stable whilemaintaining the chemical, physical and allosteric characteristics forspecific interaction with lectins or galectins known to recognizecarbohydrates. In some embodiments, the digalactoside oroligosaccharides of galactose of the invention are metabolically morestable than compounds having an O-glycosidic bond and resistant to mostgalacosidase digestion. In some embodiments, the digalactoside oroligosaccharides of galactose of the invention are metabolically morestable than compounds having an S-glycosidic bond.

Aspects of the invention relate to compounds having based on galactosidestructure with “amide” type bridge [AM] to another galactose, hydroxylcyclohexane, aromatic moiety, alkyl, aryl, amine, or amide group.

As used herein, the term “alkyl group” is meant to comprise from 1 to 12carbon atoms, for example 1 to 7 or 1 to 4 carbon atoms. In someembodiments, the alkyl group may be a straight- or a branched-chain. Insome embodiments, the alkyl group may also form a cycle comprising from3 to 7 carbon atoms, preferably 3, 4, 5, 6, or 7 carbon atoms. Thusalkyl encompasses any of methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, tert-butyl, pentyl, isopentyl, 3-methylbutyl,2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 2,2-dimethylbutyl,2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and 1-methylcyclopropyl.

As used herein, the term “alkenyl group” is meant to comprise from 2 to12, for example 2 to 7 carbon atoms. The alkenyl group comprises atleast one double bond. In some embodiments, the alkenyl groupencompasses any of vinyl, allyl, but-1-enyl, but-2-enyl,2,2-dimethylethenyl, 2,2-dimethylprop-1-enyl, pent-1-enyl, pent-2-enyl,2,3-dimethylbut-1-enyl, hex-1-enyl, hex-2-enyl, hex-3-enyl,prop-1,2-dienyl, 4-methylhex-1-enyl, cycloprop-1-enyl group, and others.

As used herein, the term “alkoxy group” relates to an alkoxy groupcontaining 1-12 carbon atoms, which may include one or more unsaturatedcarbon atoms. In some embodiments the alkoxy group contains 1 to 7 or 1to 4 carbon atoms, which may include one or more unsaturated carbonatoms. Thus the term “alkoxy group” encompasses a methoxy group, anethoxy group, a propoxy group, a isopropoxy group, a n-butoxy group, asec-butoxy group, tert-butoxy group, pentoxy group, isopentoxy group,3-methylbutoxy group, 2,2-dimethylpropoxy group, n-hexoxy group,2-methylpentoxy group, 2,2-dimethylbutoxy group 2,3-dimethylbutoxygroup, n-heptoxy group, 2-methylhexoxy group, 2,2-dimethylpentoxy group,2,3-dimethylpentoxy group, cyclopropoxy group, cyclobutoxy group,cyclopentyloxy group, cyclohexyloxy group, cycloheptyloxy group, and1-methylcyclopropyloxy group.

As used herein, the term “aryl group” is meant to comprise from 4 to 12carbon atoms. Said aryl group may be a phenyl group or a naphthyl group.The above-mentioned groups may naturally be substituted with any otherknown substituents within the art of organic chemistry. The groups mayalso be substituted with two or more of the said substituents. Examplesof substituents are halogen, alkyl, alkenyl, alkoxy, nitro, sulfo,amino, hydroxy, and carbonyl groups. Halogen substituents can be bromo,fluoro, iodo, and chloro. Alkyl groups are as defined above containing 1to 7 carbon atoms. Alkenyl are as defined above containing 2 to 7 carbonatoms, preferably 2 to 4. Alkoxy is as defined below containing 1 to 7carbon atoms, preferably 1 to 4 carbon atoms, which may contain anunsaturated carbon atom. Combinations of substituents can be presentsuch as trifluoromethyl.

As used herein, the term “heteroaryl group” is meant to comprise anyaryl group comprising from 4 to 18 carbon atoms, wherein at least oneatom of the ring is a heteroatom, i.e. not a carbon. In someembodiments, the heteroaryl group may be a pyridine, or an indole group.

The above-mentioned groups may be substituted with any other knownsubstituents within the art of organic chemistry. The groups may also besubstituted with two or more of the substituents. Examples ofsubstituents are halogen, alkoxy, nitro, sulfo, amino, hydroxy, andcarbonyl groups. Halogen substituents can be bromo, fluoro, iodo, andchloro. Alkyl groups are as defined above containing 1 to 7 carbonatoms. Alkenyl are as defined above containing 2 to 7 carbon atoms, forexample 2 to 4. Alkoxy is as defined below containing 1 to 7 carbonatoms, for example 1 to 4 carbon atoms, which may contain an unsaturatedcarbon atom. In some embodiments, the substituents can comprise

a) an alkyl group of at least 3 carbons, an alkenyl group of at least 3carbons, an alkyl group of at least 3 carbons substituted with a carboxygroup, an alkenyl group of at least 3 carbons substituted with a carboxygroup, an alkyl group of at least 3 carbons substituted with an aminogroup, an alkenyl group of at least 3 carbons substituted with an aminogroup, an alkyl group of at least 3 carbons substituted with both anamino and a carboxy group, an alkenyl group of at least 3 carbonssubstituted with both an amino and a carboxy group, and an alkyl groupsubstituted with one or more halogens. Halogens can be a fluoro, achloro, a bromo or an iodo group.

b) a phenyl group substituted with at least one carboxy group, a phenylgroup substituted with at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group,

c) a naphthyl group, a naphthyl group substituted with at least onecarboxy group, a naphthyl group substituted with at least one halogen, anaphthyl group substituted with at least one alkoxy group, a naphthylgroup substituted with at least one nitro group, a naphthyl groupsubstituted with at least one sulfo group, a naphthyl group substitutedwith at least one amino group, a naphthyl group substituted with atleast one alkylamino group, a naphthyl group substituted with at leastone dialkylamino group, a naphthyl group substituted with at least onehydroxy group, a naphthyl group substituted with at least one carbonylgroup and a naphthyl group substituted with at least one substitutedcarbonyl group; and

d) a heteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group;

e) saccharide; a substituted saccharide, D-galactose, substitutedD-galactose, C3-[1,2,3]-triazol-1-yl-substituted D-galactose, hydrogen,an alkyl group, an alkenyl group, an aryl group, a heteroaryl group, anda heterocycle and derivatives, an amino group, a substituted aminogroup, an imino group, or a substituted imino group.

Wherein NRx is selected from the group consisting of hydrogen, an alkylgroup, an alkenyl group, an aryl group, a heteroaryl group, and aheterocycle.

As used herein, the term “alkoxy group” relates to an alkoxy groupcontaining 1-7 carbon atoms, which may include one or more unsaturatedcarbon atoms. In some embodiments the alkoxy group contains 1-4 carbonatoms, which may include one or more unsaturated carbon atoms. Thus theterm “alkoxy group” encompasses a methoxy group, an ethoxy group, apropoxy group, a isopropoxy group, a n-butoxy group, a sec-butoxy group,tert-butoxy group, pentoxy group, isopentoxy group, 3-methylbutoxygroup, 2,2-dimethylpropoxy group, n-hexoxy group, 2-methylpentoxy group,2,2-dimethylbutoxy group 2,3-dimethylbutoxy group, n-heptoxy group,2-methylhexoxy group, 2,2-dimethylpentoxy group, 2,3-dimethylpentoxygroup, cyclopropoxy group, cyclobutoxy group, cyclopentyloxy group,cyclohexyloxy group, cycloheptyloxy group, and 1-methylcyclopropyloxygroup.

Monomeric Compound

In some embodiments, the compound or a pharmaceutically acceptable saltor solvate thereof has Formula 1

wherein A is independently selected from the group consisting of NRa,CRb, PRc, and amino acid,wherein M is independently selected from the group consisting of NRa,CRb, PRc, ORd, SRe amino acid, and hydrophobic hydrocarbons derivativesincluding heterocyclic substitutions of 3 or more atoms,wherein Ra is selected from the group consisting of H, H2, CH3, COOH,NH2, COMe, halogen and combinations thereof,wherein Rb is selected from the group consisting of H, H2, O, OH, CH3,COOH, NH2, COMe, halogen and combinations thereof,wherein Rc is selected from the group consisting of O2, PO2, OH, halogenand combinations thereof,wherein Rd is selected from the group consisting of H, CH3, ancombinations thereof,wherein Re is selected from the group consisting of OH, O2, S, halogenand combinations thereof,wherein B is OH, NH2, NHAc, or NH-alkyl, wherein the alkyl groupcomprises 1 to 18 Carbons,wherein W is selected from the group consisting of O, S, CH2, NH, andSe,wherein Y is selected from the group consisting of O, S, NH, CH2, Se, S,P, amino acid, and hydrophobic linear and cyclic hydrophobichydrocarbons derivatives including heterocyclic substitutions ofmolecular weight of about 50-200 D and combinations thereof,wherein R₁, R₂, and R₃ are independently selected from the groupconsisting of H, O2, CO, NH2, SO2, SO, PO2, PO, CH3, linear hydrocarbon,and cyclic hydrocarbon, andwherein the hydrocarbon is one of a) an alkyl group of at least 3carbons, an alkenyl group of at least 3 carbons, an alkyl group of atleast 3 carbons substituted with a carboxy group, an alkenyl group of atleast 3 carbons substituted with a carboxy group, an alkyl group of atleast 3 carbons substituted with an amino group, an alkenyl group of atleast 3 carbons substituted with an amino group, an alkyl group of atleast 3 carbons substituted with both an amino and a carboxy group, analkenyl group of at least 3 carbons substituted with both an amino and acarboxy group, and an alkyl group substituted with one or more halogens,b) a phenyl group substituted with at least one carboxy group, a phenylgroup substituted with at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group, c) a naphthyl group, a naphthylgroup substituted with at least one carboxy group, a naphthyl groupsubstituted with at least one halogen, a naphthyl group substituted withat least one alkoxy group, a naphthyl group substituted with at leastone nitro group, a naphthyl group substituted with at least one sulfogroup, a naphthyl group substituted With at least one amino group, anaphthyl group substituted with at least one alkylamino group, anaphthyl group substituted with at least one dialkylamino group, anaphthyl group substituted with at least one hydroxy group, a naphthylgroup substituted with at least one carbonyl group and a naphthyl groupsubstituted with at least one substituted carbonyl group, d) aheteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group, and e) asaccharide, a substituted saccharide, D-galactose, Deoxygalactose,substituted D-Galctose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose,hydrogen, an alkyl group, an alkenyl group, an aryl group, a heteroarylgroup, and a heterocycle and derivatives, an amino group, a substitutedamino group, an imino group, or a substituted imino group.

In some embodiments, wherein A-M represents a spacer of at least 2 atomscomprising an amide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, amethylether —C(—H2)-O— methylester —C(═O)—O—, carbosulfon—C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, Hydrazide —N(—H)—N(—H)—, amino acid, orcombinations thereof,

In some embodiments, the compound has the general formula (1), whereinAM-R1 is for example N′-methylamide-3,4-difluorobenzene, wherein Y—R1 istriazole-3-fluorobenzene

Dimeric Compounds

In some embodiments, the compound is adimeric-polyhydroxylated-cycloalkanes compound.

In some embodiments, the compound or a pharmaceutically acceptable saltor solvate thereof has Formula 2:

wherein A-M is representing a spacer of at least 2 atoms comprising anamide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, a methylether—C(—H2)-O— methylester —C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—,phosphate —O—P(═O)(—OH)—, diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—,carbohydrazide —C(═O)—NH—NH—, sulfonohydrazide —S(═O)2-NH—NH—, Hydrazide—N(—H)—N(—H)—, phosphonic dihydrazide —P(═O)(—NH—NH2)(NH—NH—) orcombinations thereof,wherein A is independently selected from NRa, CRb, PRc, and amino acid,wherein M is independently selected from of NRa, CRb, PRc, ORd, SReamino acid, and hydrophobic hydrocarbons derivatives includingheterocyclic substitutions of 3 or more atoms,wherein Ra is selected from the group consisting of H, H2, CH3, COOH,NH2, COMe, halogen and combinations thereof,wherein Rb is selected from the group consisting of H, H2, O, OH, CH3,COOH, NH2, COMe, halogen and combinations thereof,wherein Rc is selected from the group consisting of O2, PO2, OH, halogenand combinations thereof,wherein Rd is selected from the group consisting of H, CH3, andcombinations thereof,wherein Re is selected from the group consisting of OH, O2, S, halogenand combinations thereof,wherein B is OH, NH2, NHAc, or NH-alkyl of 1 to 18 Carbons,wherein W is selected from the group consisting of O, S, CH2, NH, or Se,Wherein X is selected from the group consisting of O, N, S, CH2, NH, andPO2,wherein Y and Z are selected from the group consisting of O, S, C, NH,CH2, Se, S, P, amino acid, and hydrophobic linear and cyclic hydrophobichydrocarbons derivatives including heterocyclic substitutions ofmolecular weight of about 50-200 D and combinations thereof,wherein R1, R2, R3, are independently selected from the group consistingof CO, O2, SO2, SO, PO2, PO, CH, Hydrogen, hydrophobic linearhydrocarbon, and hydrophobic cyclic hydrocarbon, wherein the hydrocarbonis one of:a) an alkyl group of at least 3 carbons, an alkenyl group of at least 3carbons, an alkyl group of at least 3 carbons substituted with a carboxygroup, an alkenyl group of at least 3 carbons substituted with a carboxygroup, an alkyl group of at least 3 carbons substituted with an aminogroup, an alkenyl group of at least 3 carbons substituted With an aminogroup, an alkyl group of at least 3 carbons substituted with both anamino and a carboxy group, an alkenyl group of at least 3 carbonssubstituted with both an amino and a carboxy group, and an alkyl groupsubstituted with one or more halogens,b) a phenyl group substituted with at least one car boxy group, a phenylgroup substituted With at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group,c) a naphthyl group, a naphthyl group substituted with at least onecarboxy group, a naphthyl group substituted with at least one halogen, anaphthyl group substituted with at least one alkoxy group, a naphthylgroup substituted with at least one nitro group, a naphthyl groupsubstituted with at least one sulfo group, a naphthyl group substitutedWith at least one amino group, a naphthyl group substituted with atleast one alkylamino group, a naphthyl group substituted with at leastone dialkylamino group, a naphthyl group substituted with at least onehydroxy group, a naphthyl group substituted with at least one carbonylgroup and a naphthyl group substituted with at least one substitutedcarbonyl group; andd) a heteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group.e) saccharide, a substituted saccharide, D-galactose, substitutedD-galactose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose, hydrogen,an alkyl group, an alkenyl group, an aryl group, a heteroaryl group, anda heterocycle and derivatives; an amino group, a substituted aminogroup, an imino group, or a substituted imino group.

In some embodiments, the compound has the general formulas below

Wherein A-M is an amide, Sulfate, sulfonamide, a carbon ester and/orincludes an aryl derivative like AM-Benzene-AM structure (Example 14,Scheme 6).Wherein W is selected from the group consisting of O, N, S, CH2, NH, andSe;Wherein Y and Z are selected from the group consisting of O, S, C, NH,CH2, NR, Se, or Amino acid.Wherein R1, R2, R3, and R4 (Rx) are independently selected from thegroup consisting of CO, SO2, SO, PO2, PO, CH, Hydrogen, Hydrophobiclinear and cyclic including Heterocyclic substitutions of molecularweight of 50-200 D including, but not limited to:a) an alkyl group of at least 3 carbons, an alkenyl group of at least 3carbons, an alkyl group of at least 3 carbons substituted with a carboxygroup, an alkenyl group of at least 3 carbons substituted with a carboxygroup, an alkyl group of at least 3 carbons substituted with an aminogroup, an alkenyl group of at least 3 carbons substituted with an aminogroup, an alkyl group of at least 3 carbons substituted with both anamino and a carboxy group, an alkenyl group of at least 3 carbonssubstituted with both an amino and a carboxy group, and an alkyl groupsubstituted with one or more halogens;b) a phenyl group substituted with at least one car boxy group, a phenylgroup substituted With at least one halogen, a phenyl group substitutedwith at least one alkoxy group, a phenyl group substituted with at leastone nitro group, a phenyl group substituted with at least one sulfogroup, a phenyl group substituted with at least one amino group, aphenyl group substituted with at least one alkylamino group, a phenylgroup substituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group,c) a naphthyl group, a naphthyl group substituted with at least onecarboxy group, a naphthyl group substituted with at least one halogen, anaphthyl group substituted with at least one alkoxy group, a naphthylgroup substituted with at least one nitro group, a naphthyl groupsubstituted with at least one sulfo group, a naphthyl group substitutedWith at least one amino group, a naphthyl group substituted with atleast one alkylamino group, a naphthyl group substituted with at leastone dialkylamino group, a naphthyl group substituted with at least onehydroxy group, a naphthyl group substituted with at least one carbonylgroup and a naphthyl group substituted with at least one substitutedcarbonyl group; andd) a heteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group.e) saccharide, a substituted saccharide, D-galactose, substitutedD-galactose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose, hydrogen,an alkyl group, an alkenyl group, an aryl group, a heteroaryl group, anda heterocycle and derivatives; an amino group, a substituted aminogroup, an imino group, or a substituted imino group.Rx is selected from the group consisting of hydrogen, an alkyl group, analkenyl group, an aryl group, a heteroaryl group, and a heterocycle.

As used herein, the term “alkyl group” relates to an alkyl groupcontaining 1-7 carbon atoms, which may include one or more unsaturatedcarbon atoms. In some embodiments the alkyl group contains 1-4 carbonatoms, which may include one or more unsaturated carbon atoms. Thecarbon atoms in the alkyl group may form a straight or branched chain.The carbon atoms in said alkyl group may also form a cycle containing 3,4, 5, 6, or 7 carbon atoms. Thus, the term “alkyl group” used hereinencompasses methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl,tert-butyl, pentyl, isopentyl, 3-methylbutyl, 2,2-dimethylpropyl,n-hexyl, 2-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, n-heptyl,2-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and1-methylcyclopropyl.

Without being bound to the theory, the galactose-Amide or sulfur basedlinker compounds described herein have an enhanced stability as itsstructure is less prone to hydrolysis (metabolism) and oxidation e.g.aromatic ring without substitutions, Carbon-Oxygen systems,Carbone-Nitrogen system etc.

Synthetic Route

The compounds of this invention may be prepared by the following generalmethods and procedures. It should be appreciated that where typical orpreferred process conditions (e.g. reaction temperatures, times, molarratios of reactants, solvents, pressures, pH etc) are given, otherprocess conditions may also be used unless otherwise stated. Optimumreaction conditions may vary with the particular reactants, solventsused and pH etc., but such conditions can be determined by one skilledin the art by routine optimization procedures.

In some embodiments, the compounds were synthetized using the syntheticroutes as given in Example 14 and shown in FIG. 4 .

For example, compound G631 (a Galactosulfonamide, GTJC-026) was preparedas shown in Example 14 Scheme 11 (as shown FIG. 4 ).

In some embodiments, the di-galactoamide compounds were synthetizedusing the synthetic routes as given in Example 14 Scheme 6 and shown inFIG. 4 .

For example, compound G637 (a di-galactoamide with arylamide linkage,GTJC-013-12) was prepared as shown in Example 14 Scheme 6 and shown inFIG. 4 .

Pharmaceutical Compositions

Aspects of the invention relate to the use of the compounds describedherein for the manufacture of medicaments.

Aspects of the invention relate to pharmaceutical compositionscomprising one or more of the compounds described herein. In someembodiments, the pharmaceutical compositions comprise one or more of thefollowing: pharmaceutically acceptable adjuvant, diluent, excipient, andcarrier.

In some embodiments, the pharmaceutical composition comprising acompound described herein may be adapted for oral, intravenous, topical,intraperitoneal, nasal, buccal, sublingual, or subcutaneousadministration, or for administration via the respiratory tract in theform of, for example, an aerosol or an air-suspended fine powder, or,for administration via the eye, intra-ocularly, intravitreally orcorneally.

T In some embodiments, the pharmaceutical composition comprising acompound described herein may be in the form of, for example, tablets,capsules, powders, solutions for injection, solutions for spraying,ointments, transdermal patches or suppositories,

Methods of Treatment

Some aspects of the invention relate to the use of the compoundsdescribed herein or the composition described herein for us in thetreatment of a disorder relating to the binding of a galectin to aligand. In some embodiments, galectin is galectin-3.

Some aspects of the invention relate to the method of treating variousdisorders relating to the binding of a galectin to a ligand. In someembodiments, the methods comprise administering in a subject in needthereof a therapeutically effective amount of at least one compounddescribed herein. In some embodiments, the subject in need thereof is ahuman having high levels of galectin-3. Levels of galectin, for examplegalectin-3 can be quantified using any methods known in the art.

In some embodiments, the disorder is an inflammatory disorder, forexample inflammatory bowel disease, Crohn's disease, multiple sclerosis,Systemic Lupus Erythematosus, ulcerative colitis.

In some embodiments, the disorder is fibrosis, for example liverfibrosis, pulmonary fibrosis, kidney fibrosis, heart fibrosis orfibrosis of any organ compromising the normal function of the organ.

In some embodiments, the disorder is cancer.

In some embodiments, the disorder is an autoimmune disease such asrheumatoid arthritis and multiple sclerosis.

In some embodiments, the disorder is heart disease or heart failure.

In some embodiments, the disorder is a metabolic disorder, for examplediabetes.

In some embodiments, the disorder relating is pathological angiogenesis,such as ocular angiogenesis, disease or conditions associated withocular angiogenesis and cancer.

In some embodiments, the compounds of the invention comprises pyranosyland/or furanosyl structures conjugated through an amide or sulfonamidetype linkage to organic substitutions and are designated as“GalactoAmides” and/or “GalactoSulfonamides” with the general structuresR′-Gal-AM-R″ where the “AM” is an “amide” or “sulfonamide” type linkagewith R′ and R″ are organic substituents.

In some embodiments, the compound comprises of functional galactose likepyranosyl and/or furanosyl structures linked through the anomeric carbonof the pyranosyl and/or furanosyl by “AM” type linkage to an organicsubstituent. As used herein the “AM” linkage is not limited to simpleamide and can be any of the following linkages: N′-methylamide,sulfonamide, C-amide, O-Succinimide, Acetohydrazide, bemethly amide,N-ethylbenzene-amide, N-ethylamide, N-methoxypropane-amide,N-methoxypropanol-amide, methyl-sulfur or sulfon linker or anycombinations of the foregoing.

In some embodiments, the organic substituents are specific aromaticsubstitutions linked to the galactose core or the “AM” linker of theanomeric carbon of the pyranosyl and/or furanosyl structures. Sucharomatic substitutions can enhance the interaction of the compound withamino acid residues (e.g. Arginine, Tryptophan, Histidine, Glutamic acidetc. . . . ) composing the carbohydrate-recognition-domains (CRD) of thelectins or with amino acid residues in the CRD neighborhood and thusstrengthen the association and binding specificity.

In some embodiments, the organic substituents comprises monosaccharides,disaccharides, oligosaccharides or a heteroglycoside such as iminosugaror thiosugar carbohydrates with nitrogen or sulfur atoms replacing theOxygen and bound to the “amide” linker on the anomeric carbon of thegalactose core.

In some embodiments, the compound is a symmetric digalactoside, whereinthe two galactosides are bound by an “amide” linker. Yet in otherembodiments, the compound can be comprised of asymmetric carbohydrates.For example, each of the galactoside can have different aromatic oraliphatic substitutions or heterotatoms derivatives of the galactosewhere the C5 oxygen is replaced with S (5-Thio-D-galactose) or N(5-imino-D-galactose).

Without being bound to the theory, it is believed that the compoundscontaining the “Amide” based linker containing molecules render thecompounds metabolically stable while maintaining the chemical, physicaland allosteric characteristics for specific interaction with lectins orgalectins known to recognize carbohydrates. In some embodiments, theGalactoAmide and the GalactoSulfonamide of the invention aremetabolically more stable than compounds having an O-glycosidic bond.

Aspect the invention relates to a compound or a pharmaceuticallyacceptable salt or solvate thereof:

Wherein X is NH, NCH2, SNH, SO2, CH2, COH, Se, or amino acid,Wherein Z is independently selected from a linkage consisting of C, NH,O, S, SO2, COH, Se to create the “AM” amide type linkage, e.g. amide,N′-methylamide, Sulfonamide, carbosulfon, Sulfonate, acetohydrazidelinkage to the substitutions R2 and R3,Wherein W is selected from the group consisting of O, N, S, CH2, NH, andSe;Wherein Y is selected from the group consisting of O, S, C, NH, CH2, Se,and amino acid;Wherein R₁, R₂, R₃, and R₄ are independently selected from the groupconsisting of H, CO, SO2, SO, PO2, PO, CH, and hydrophobic linear andcyclic hydrocarbons including heterocyclic substitutions of molecularweight of about 50-200 D.

In some embodiments, the AM linkage comprise Se or Se—Se bond and one ormore atoms such as C or N. In some embodiments, Se can be directlylinked to the anomeric Carbon and linked to C, N or O. In someembodiments, Se can be in the second position and C or N are linked tothe anomeric carbon.

In some embodiments, the hydrophobic linear and cyclic hydrocarbons cancomprise one of: a) an alkyl group of at least 4 carbons, an alkenylgroup of at least 4 carbons, an alkyl group of at least 4 carbonssubstituted with a carboxy group, an alkenyl group of at least 4 carbonssubstituted with a carboxy group, an alkyl group of at least 4 carbonssubstituted with an amino group, an alkenyl group of at least 4 carbonssubstituted with an amino group, an alkyl group of at least 4 carbonssubstituted with both an amino and a carboxy group, an alkenyl group ofat least 4 carbons substituted with both an amino and a carboxy group,and an alkyl group substituted with one or more halogens, b) a phenylgroup substituted with at least one carboxy group, a phenyl groupsubstituted with at least one halogen, a phenyl group substituted withat least one alkoxy group, a phenyl group substituted with at least onenitro group, a phenyl group substituted with at least one sulfo group, aphenyl group substituted with at least one amino group, a phenyl groupsubstituted with at least one alkylamino group, a phenyl groupsubstituted with at least one dialkylamino group, a phenyl groupsubstituted with at least one hydroxy group, a phenyl group substitutedwith at least one carbonyl group and a phenyl group substituted with atleast one substituted carbonyl group, c) a naphthyl group, a naphthylgroup substituted with at least one carboxy group, a naphthyl groupsubstituted with at least one halogen, a naphthyl group substituted withat least one alkoxy group, a naphthyl group substituted with at leastone nitro group, a naphthyl group substituted with at least one sulfogroup, a naphthyl group substituted With at least one amino group, anaphthyl group substituted with at least one alkylamino group, anaphthyl group substituted with at least one dialkylamino group, anaphthyl group substituted with at least one hydroxy group, a naphthylgroup substituted with at least one carbonyl group and a naphthyl groupsubstituted with at least one substituted carbonyl group, d) aheteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group, and e) asaccharide, a substituted saccharide, D-galactose, substitutedD-galactose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose, hydrogen,an alkyl group, an alkenyl group, an aryl group, a heteroaryl group, anda heterocycle and derivatives, an amino group, a substituted aminogroup, an imino group, or a substituted imino group.

EXAMPLES Example 1: Compound Inhibition of Galectin Binding toPhysiologic Ligands

Galectin proteins, including but not limited to galectin-3 andgalectin-1, have multiple biologically relevant binding ligands inmammalian species, including but not limited to rodents, primates, andhumans. Galectins are carbohydrate-binding proteins that bind toglycoproteins with β-galactoside-containing sugars. The result ofbinding of galectin proteins to these ligands results in a plethora ofbiological effects in and on cells and in tissues and whole organismsincluding regulating cell survival and signaling, influencing cellgrowth and chemotaxis, interfering with cytokine secretion, mediatingcell-cell and cell-matrix interactions or influencing tumor progressionand metastasis. Additionally, changes in normal expression of galectinproteins are responsible for pathological effects in multiple diseases,including but not limited to inflammatory, fibrotic and neoplasticdiseases. (See FIGS. 8B, 9 )

To screen galectin-3 functional activity and neutralizing effects ofcompounds of the invention, specific monoclonal antibodies have beenselected and analytical assay using an ELISA format was developed asillustrate in FIG. 5A. The inhibitory comparison of this antibodiesbased ELISA with the inhibition of Integrin amB2 to galectin-3 had goodregression factor of better than 0.95 as presented in FIG. 7A and FIG.7B.

Compounds described herein are designed to bind to the carbohydraterecognition domain of galectin proteins, including but not limited togalectin-3, and disrupt interactions with biologically relevant ligands.They are intended to inhibit the function of galectin proteins that maybe involved in pathological processes at normal levels of expression orin situations where they are increased over physiological levels.

Some of the ligands for galectin proteins that are important in normalcellular function and pathology in disease include, but are not limitedto, TIM-3 (T cell immunoglobulin mucin-3)), CD8, T cell receptor,integrins, galectin-3 binding protein, TGF-β receptor, laminins,fibronectins, BCR (B cell receptor, CTLA-4 (cytotoxicT-lymphocyte-associated protein-4), EGFR (Epidermal growth factorreceptor), FGFR (fibroblast growth factor receptor), GLUT-2 (glucosetransporter-2), IGFR (insulin-like growth factor receptor), variousinterleukins, LPG (lipophosphoglycan), MHC (major histocompatibilitycomplex), PDGFR (platelet-derived growth factor receptor), TCR (T cellreceptor), TGF-β (transforming growth factor-β), TGFPR (transforminggrowth factor-β receptor, CD98, Mac3 antigen (Lysosome-associatedmembrane protein 2 (LAMP2) also known as CD107b (Cluster ofDifferentiation 107b)).

Experiments have been performed to evaluate the physical interaction ofgalectin proteins with these various biological ligands mediatingcellular functions. The experiments were designed to evaluate theinteraction between various galectin-3 ligands and determine whethercompounds described herein are able to inhibit these interactions, asanalytical assays format shown in FIG. 5B.

Using these assays format, the compounds described herein inhibit theinteraction of galectin proteins with their ligands, including but notlimited to various integrin molecules (αVβ3, αVβ6, αMβ2, α2ρ3, andothers) with IC50's in the range of about 5 ηM to about 40 μM. In someembodiments, the IC50 is about from 5 nM to about 20 nM. In someembodiments, the IC50 is from about 5 nM to about 100 nM. In someembodiments, the IC50 is from about 10 nM to about 100 nM. In someembodiments, the IC50 is from about 50 nM to about 5 μM. In someembodiments, the IC50 is from about 0.5 μM to about 10 μM. In someembodiments, the IC50 is from about 5 μM to about 40 μM as listed inFIG. 7A and FIG. 7B). Further examples of the inhibition effect ofexemplary compounds of the invention of galectin-3 interaction withIntegrin amB2 are shown in FIG. 8B and of the inhibition of galectin-3with integrin aVM6 are shown in FIG. 9 .

Example 2: Compound Inhibition of Galectin Binding to Labeled Probes

Fluorescein-labeled probes have been developed which bind to galectin-3and other galectin proteins and these probes have been used to establishassays that measure the binding affinity of ligands for the galectinproteins using Fluorescence Polarization (Sorme P, et al. Anal Biochem.2004 Nov. 1,334(1):36-47).

Compounds described herein avidly bind to galectin-3, as well as othergalectin proteins, using this assay format (FIG. 6B) and displace theprobe with high affinity, with IC50's (concentration at 50% inhibition)of between about 5 ηM to about 40 μM. In some embodiments, the IC50 isabout from 5 nM to about 20 nM. In some embodiments, the IC50 is fromabout 5 nM to about 100 nM. In some embodiments, the IC50 is from about10 nM to about 100 nM. In some embodiments, the IC50 is from about 50 nMto about 5 μM. In some embodiments, the IC50 is from about 0.5 μM toabout 10 μM. In some embodiments, the IC50 is from about 5 μM to about40 μM.

Exemplary compounds of the invention were synthesized (FIG. 4 ) andshowed an inhibitory activity in the Fluorescent polarization assay(FIG. 8A).

Example 3: Compound Inhibition of Galectin Binding Using FRET Assay

FRET assay (fluorescent resonance energy transfer) assays were developedfor evaluating the interaction of galectin proteins, including but notlimited to galectin-3, with a model fluorescent-labeled probe (see FIG.6A). Using this assay, compounds described herein avidly bind togalectin-3, as well as other galectin proteins, using this assay anddisplace the probe with high affinity, with IC₅₀'s (concentration at 50%inhibition) of between about 5 ηM to about 40 μM. In some embodiments,the IC50 is about from 5 nM to about 20 nM. In some embodiments, theIC50 is from about 5 nM to about 100 nM. In some embodiments, the IC50is from about 10 nM to about 100 nM. In some embodiments, the IC50 isfrom about 50 nM to about 5 μM. In some embodiments, the IC50 is fromabout 0.5 μM to about 10 μM. In some embodiments, the IC50 is from about5 μM to about 40 μM.

Example 4: Compound Binding to Amino Acid Residues in Galectin Proteins

Heteronuclear NMR spectroscopy is used to evaluate the interaction ofcompounds described herein with galectin molecules, including but notlimited to galectin-3, to assess the interaction residues on thegalectin-3 molecule.

Uniformly ¹⁵N-labeled Gal-3 is expressed in BL21 (DE3) competent cells(Novagen), grown in minimal media, purified over a lactose affinitycolumn, and fractionated on a gel filtration column, as describedpreviously for production of Gal-1 (Nesmelova I V, Pang M, Baum L G,Mayo K H. 1H, 13C, and 15N backbone and side-chain chemical shiftassignments for the 29 kDa human galectin-1 protein dimer. Biomol NMRAssign 2008 December; 2(2):203-205).

Uniformly ¹⁵N-labeled Gal-3 is dissolved at a concentration of 2 mg/mlin 20 mM potassium phosphate buffer at pH 7.0, made up using a 95%H₂O/5% D₂O mixture. ¹H-¹⁵N HSQC NMR experiments are used to investigatebinding of a series of compounds described herein. ¹H and ¹⁵N resonanceassignments for recombinant human Gal-3 were previously reported (IppelH, et al. (1)H, (13)C, and (15)N backbone and side-chain chemical shiftassignments for the 36 proline-containing, full length 29 kDa humanchimera-type galectin-3. Biomol NMR Assign 2015 April; 9(1):59-63.).

NMR experiments are carried out at 30° C. on Bruker 600 MHz, 700 MHz or850 MHz spectrometers equipped with H/C/N triple-resonance probes andx/y/z triple-axis pulse field gradient units. A gradientsensitivity-enhanced version of two-dimensional ¹H-¹⁵N HSQC is appliedwith 256 (t1)×2048 (t2) complex data points in nitrogen and protondimensions, respectively. Raw data are converted and processed by usingNMRPipe and were analyzed by using NMRview.

These experiments show differences between compounds described hereinand galactose in the binding residues in the carbohydrate binding domainof galectin-3.

Example 5: Cellular Activity of Cytokine Activity Related to GalectinBinding Inhibition

Example 1 describes the ability of compounds of this application toinhibit the binding of physiologic ligands to galectin molecules. In theexperiments of this example, the functional implications of thosebinding interactions are evaluated.

One of the interactions with galectin-3 that is inhibited by thecompounds described herein was TGF-β receptor. Therefore, experimentsare done to evaluate the effect of compounds on TGR-β receptor activityin cell lines. Various TGF-β responsive cell lines, including but notlimited to LX-2 and THP-1 cells, are treated with TGF-β and response ofthe cells is measured by looking at activation of second messengersystems, including but not limited to phosphorylation of variousintracellular SMAD proteins. After establishing that TGF-βactivates thesecond messenger systems in the various cell lines, the cells aretreated with compounds described herein. This experiments show thatthese compounds inhibit TGF-β signaling pathways, confirming that thebinding interaction inhibition described in Example 1 has aphysiological role in cellular models.

Cellular assays are also performed to evaluate the physiologicalsignificance of inhibiting the interaction of galectin-3 with variousintegrin molecules. Cell-cell interaction studies are performed usingmonocytes binding to vascular endothelial cells, as well as other celllines. Treatment of cells with compounds described herein is found toinhibit these integrin-dependent interactions, confirming that thebinding interaction inhibition described in Example 1 has aphysiological role in cellular models.

Cellular motility assays are performed to evaluate the physiologicalsignificance of inhibiting the interaction of galectin-3 with variousintegrin and other cell surface molecules defined in Example 1. Cellularstudies are performed using multiple cell lines in a semi-permeablemembrane separated well apparatus. Treatment of cells with compoundsdescribed herein is found to inhibit cellular motility, confirming thatthe binding interaction inhibition described in Example 1 has aphysiological role in cellular models.

Example 6: In-Vitro Inflammatory Model (a Monocyte Based Assay)

A model of macrophage polarization is set up, starting from THP-1monocytes culture which is differentiated into inflammatory macrophagesusing PMA (Phorbol 12-myristate 13-acetate) for 2-4 days. Oncedifferentiated (M0 macrophages), the macrophages are induced with LPS orLPS and IFN-gamma for macrophage activation (M1) to inflammatory stagefor 1-3 days. Array of cytokines and chemokines are analyzed to confirmthe polarization of THP-1-derived macrophages to inflammatory stage. Theimpact of the anti-galectin-3 compounds on macrophage polarization isassessed first by monitoring cell viability using a colorimetric method(using a tetrazolium reagent) to determine the number of viable cells inproliferation or cytotoxicity assays (Promega, The CellTiter 96® AQueousOne Solution Cell Proliferation Assay which contains a novel tetrazoliumcompound[3-(4,5-dimethyl-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt; MTS] and an electron coupling reagent (phenazineethosulfate; PES)) and inflammatory stage evaluated by a quantitativelymeasure of the chemokine Monocyte Chemoattractant Protein-1(MCP-1/CCL2), a key protein that regulates migration and infiltration ofmonocytes/macrophages in cellular process of inflammation. Follow-uptesting for the expression and secretion of other cytokines andchemokines are done for leading active compounds. Results are expressedin percentage reduction of MCP-1 as shown in FIG. 10A for compoundsaccording to some embodiments.

Example of method steps:

1) THP-1 cells were cultured in media containing Gentamicin2) THP-1 cells are transfer to wells in a 96 well plate 2,000 cells/wellfor 2 days incubation in assay media containing 5-50 ng/ml PMA3) Serial dilution of test compounds is made in LPS (1-10 ng/ml)containing media4) To each well 100 ml of compounds/LPS solution is added to a finalassay volume of each well of 200 ml which contains also Gentamicin and5-20 ng/ml PMA5) Cells are incubated up to 8 days.6) Every other day samples of 20-60 ul are removed for biomarker assays7) At termination 15 ml of Promega Substrate CellTiter 96 Aqueous OneSolution is prepared and added to each well to monitor cytotoxicity (at490 nm)8) For cellular biomarkers evaluation the cells are washed 1×PBS andextracted with 200 ul of Lysis buffer for 1 hour. Extract is spinneddown 10 minutes and 120 ul sample is removed from top. All samples arekept at −70 C until testing. (See FIG. 10 )

Example 7: Cell Culture Fibrogenesis Model

Experiments are performed with fibrogenic stellate cell cultures,including but not limited to LX-2 cells, to evaluate the cellular effectof compounds herein. LX-2 cells are activated in culture using serumdeprived media and media spiked with different percentages of THP-1 cellconditioned media. Activation of LX-2 cells is monitored by various welldefined markers, including but not limited to TIMP-1. Demonstrable LX-2cell activation is evident by 24 hours after treatment. The treatment ofcells with compounds described herein is found to inhibit activation,confirming a physiological role in cellular models.

Example 8: In Vivo Animal Models of Liver Fibrosis NASH Mouse FibrosisModel

The NASH model uses male newborn mice [C57BL/6J mice]. The disease isinduced by a single subcutaneous injection of streptozotocin (Sigma, St.Louis, Mo.) solution 2 days after birth which induced diabetes. Afterfour weeks of age a high fat diet (HFD, 57% of kcal from fat) isintroduced for 12 and up to 16 weeks as demonstrate in the time lineenclosed graph. Vehicle and test substances at the various doses areadministered orally or SQ or intravenously weekly and calculated asmg/kg body weight. Animal care follows protocols accordance withaccepted Guidelines for Animal Use. Animals are fasted for 3 hoursbefore sacrifice which is performed by exsanguination through directcardiac puncture under ether anesthesia.

Randomization of mice into treatment groups is done prior to treatmentbased on the plasma ALT levels and body weight. At minimum 3 treatmentgroups are in a study.

Group 1: Twelve normal mice are fed with a normal diet ad libitumwithout any treatment,

Group 2: Twelve NASH mice are intravenously administered vehicle (0.9%sodium chloride) once weekly from 6 to 12 weeks of age

Group 3: Twelve NASH mice are intravenously administered test article invehicle (0.9% sodium chloride) once weekly from 6 to 12 weeks of age

Mice are sacrificed for the following 4 weeks of treatment

The seleno-galactoside compounds described herein reduce live fibrosisas measured by collagen 10% to 80% versus the vehicle control or toalmost normal collagen levels as established in group 1.

General Biochemical Tests:

Diabetic fast glucose is measured in whole blood samples using forexample G Checker (Sanko Junyaku Co. Ltd., Japan).

Liver functions are evaluated in Plasma for levels of AST, ALT, totalbilirubin, creatinine, and TG are measured by example FUJI DRY CHEM 7000(Fuji Film, Japan).

Liver biochemistry: To quantify liver hydroxyproline content, aquantitative assessment of collagen content, frozen liver samples (40-70mg) are processed by a standard alkaline-acid hydrolysis method andhydroxyproline content is normalized to total liver proteins.

Total liver lipid-extracts are obtained from caudate lobes by Folch'smethod and liver TG levels are measured using the Triglyceride E-test(Wako, Japan).

Histopathological and immunohistochemical analyses liver sections arecut from paraffin blocks of liver tissue prefixed in Bouin's solutionand stained with Lillie-Mayer's Hematoxylin (Muto Pure Chemicals, Japan)and eosin solution (Wako, Japan).

To visualize collagen deposition, Bouin's fixed liver sections arestained using picro-Sirius red solution (Waldeck GmbH & Co. KG,Germany). NAFLD Activity score (NAS) is also calculated according toestablished criteria.

Immunohistochemistry for SMA, F4/80, Galectin-3, CD36 and iNOS can beestimated from each positive area as indication for the extent ofinflammation and fibrosis.

Rat Fibrosis/Cirrhosis Model (TAA Model):

These experiments use male Sprague-Dawley rats between 160 and 280 gobtained from animal research facility (Jackson Laboratory) which aremaintained according to the Guide for the Care and Use of LaboratoryAnimals (Institute of Laboratory Animal Resources, 1996, Nat. Acad.Press) and Institutional Animal Care and Use committee (IACUC). At theend of experiments, animals are euthanized under phenobarbitalanesthesia.

After an acclimation period of two weeks, an eight week induction periodis initiated, in which all rats are subjected to intraperitoneal (IP)injections Thioacetamide (TAA, Sigma Chemical Co., St. Louis, Mo., USA)of sterile solutions of dissolved in 0.9% saline, administered by IPinjection twice or trice weekly with initial week dosage of 450mg/kg/wk, followed by seven weeks regimen of 400 mg/kg/wk body weight.To assess for the progression of fibrosis two rats are euthanized atweeks 4 and 8, and the liver examined histologically. To developcirrhosis animals are administered TAA intraperitoneally (IP) up to11-12 weeks, for fibrosis 8 weeks are enough. Treatment is for 4 weeksbeginning in week 8, vehicle control group is administered 0.9% NaClintraperitoneally twice weekly for four weeks. Experimental testarticles are given intraperitoneally twice or once a week beginning inweek 8 or 11 for fibrosis or cirrhosis respectively. At the end of thetreatment period, rats are placed under anesthesia using isofluoranebetween 1-5% through inhalation and a laparotomy is performed. At thetime of sacrifice, portal pressure is measured using a 16 Gangiocatheter introduced into the portal vein to measure the height of awater column. The liver is removed, weighed, and pieces from the largestlobes are used for further analysis. The spleen is also removed andweighed before being discarded.

Representative histology of Sirius red stained liver sections fromexperiment shows a 20% reduction in mean collagen which is statisticalacceptable for anti-fibrosis effect. Strands of bridging fibrosisindicate advance fibrosis stage (these are strands of collagen fibers).

Biochemical Tests:

As in the NASH model various diagnostic tests are done to evaluate theextend of liver damage due to the fibrosis:

Liver functions are evaluated in Plasma for levels of AST, ALT, totalbilirubin, creatinine, and TG are measured by example FUJI DRY CHEM 7000(Fuji Film, Japan).

Liver biochemistry: To quantify liver hydroxyproline content, aquantitative assessment of collagen content, frozen liver samples (40-70mg) are processed by a standard alkaline-acid hydrolysis method andhydroxyproline content is normalized to total liver proteins.

Total liver lipid-extracts are obtained from caudate lobes by Folch'smethod and liver TG levels are measured using the Triglyceride E-test(Wako, Japan).

Histopathological and immunohistochemical analyses liver sections arecut from paraffin blocks of liver tissue prefixed in Bouin's solutionand stained with Lillie-Mayer's Hematoxylin (Muto Pure Chemicals, Japan)and eosin solution (Wako, Japan).

To visualize collagen deposition, Bouin's fixed liver sections arestained using picro-Sirius red solution (Waldeck GmbH & Co. KG,Germany). NAFLD Activity score (NAS) is also calculated according toestablished criteria.

Immunohistochemistry for SMA, F4/80, Galectin-3, CD36 and iNOS can beestimated from each positive area as indication for the extent ofinflammation and fibrosis.

Bile Duct Models of Liver Fibrosis

These experiments are done to evaluate the efficacy of the compoundsdescribed herein on the fibrosis of the liver following bile ductligation or treatment with drugs that cause biliary fibrosis. Animalstreated with the compounds herein described show that liver fibrosis wasreduced in comparison to vehicle controls.

Example 9: In Vivo Animal Models of Lung Fibrosis

These experiments are done to evaluate the efficacy of the compoundsdescribed herein on the prevention of bleomycin-induced pulmonaryfibrosis. An untreated control group with intratracheal saline infusionconsists of 10 mice. Bleomycin is administered by slow intratrachealinfusion into the lungs of other groups on Day 0. On Days −1, 2, 6, 9,13, 16 and 20, mice are dosed (iv, ip, subcut, or oral) once daily withvehicle or various doses of compounds described herein (iv, ip, subcut,or oral). Animals are weighed and evaluated for respiratory distressdaily. On Day 21, all animals are euthanized and the wet weight of lungsis measured. Upon sacrifice, blood is collected via retro-orbital bleedfor preparation of serum. The right lobe of the lung is snap frozen forsubsequent hydroxyproline analysis while the left is insufflated andfixed in 10% formalin for histological analysis. The formalin-fixed lungis processed for routine histological evaluation.

Example 10: In Vivo Animal Models of Kidney Fibrosis

These experiments are done to evaluate the efficacy of the compoundsdescribed herein on the fibrosis of the kidney using models ofunilateral ureteral ligation and diabetic nephropathy. Animals treatedwith various compounds herein show that kidney fibrosis is reduced incomparison to vehicle controls.

Example 11: In Vivo Animal Models of Cardiovascular Fibrosis

These experiments are done to evaluate the efficacy of the compoundsdescribed herein on the fibrosis of the heart and vessels using modelsof heart failure, atrial fibrillation, pulmonary hypertension, andatherosclerosis. Animals treated with various compounds herein show thatcardiovascular fibrosis was reduced in comparison to vehicle controls.

Example 12: VEGF-A-Induced Angiogenesis

Vascular endothelial growth factors (VEGFs) signaling though VEGFreceptor-2 (VEGFR-2) is the primary angiogenic pathway. Galectinproteins are important for the signaling pathway. Compounds describedherein are able to inhibit neovascularization of mouse cornea inresponse to injury.

Example 13: Evaluation of Compound Absorption, Distribution, Metabolism,and Elimination

Compounds described herein are evaluated for physicochemical properties,including but not limited to solubility (Thermodynamic and Kineticmethod), various pH changes, solubility in biorelevant medium (FaSSIF,FaSSGF, FeSSIF), Log D (Octanol/water and Cyclohexane/water), chemicalstability in plasma, and blood partitioning.

Compounds described herein are evaluated for in vitro permeabilityproperties, including but not limited to PAMPA (parallel artificialmembrane permeability assay), Caco-2, and MDCK (wild type)

Compounds described herein are evaluated for animal pharmacokineticproperties, including but not limited to pharmacokinetics by variousroutes viz., oral, intravenous, intraperitoneal, subcutaneous in mice(Swiss Albino, C57, Balb/C), rats (Wistar, Sprague Dawley), rabbits (NewZealand white), dogs (Beagle), Cynomolgus monkeys, etc., tissuedistribution, brain to plasma ratio, biliary excretion, and massbalance.

Compounds described herein are evaluated for protein binding, includingbut not limited to plasma protein binding (ultra Filtration andEquilibrium Dialysis) and microsomal protein binding.

Compounds described herein are evaluated for in vitro metabolism,including but not limited to cytochrome P450 inhibition, cytochrome P450time dependent inhibition, metabolic stability, liver microsomemetabolism, S-9 fraction metabolism, effect on cryopreserved hepatocyte,plasma stability, and GSH trapping.

Compounds described herein are evaluated for metabolite identification,including but not limited to identification in vitro (microsomes, S9 andhepatocytes) and in vivo samples.

Example 14: Synthesis of GalactoAmide and GalactoSulfonamides Compounds

Non-limiting examples of compounds according to some embodiments areshown in Table 1 and FIG. 4 .

Step 1(3aR,5R,6aS)-5-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dinethyldihydrofuro[2,3-d][1,3]dioxol-6(5H)-one

To a stirred solution of(3aR,5S,6S,6aR)-5-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol(1000 g, 3846 mmol) in DCM (8000 mL) was added Ac2O (3.9 eq) followed byPDC (1.5 eq) portion wise at room temperature over a period of 2 h. Thereaction mixture was reflux for 3 h. After completion, the crude productwas passed through a SiO2 column (60-120 mesh, 15 kg) and eluted withethyl acetate (40 L). The solvent was evaporated to afford the titlecompound as a sticky yellow liquid (580 g, 58%). 1H NMR (400 MHz;CDCl3): □ 6.13 (d, J=4.4 Hz, 1H), 4.35-4.42 (m, 3H), 4.01-4.07 (m, 2H),1.54 (s, 3H), 1.44 (s, 3H), 1.36 (s, 3H), 1.31 (s, 3H).

Step 2

(3aR,6aR)-5-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyl-3a,6a-dihydrofuro[2,3-d][1,3]dioxol-6-yl acetate: To a stirred solution of(3aR,5R,6aS)-5-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyldihydrofuro[2,3-d][1,3]dioxol-6(5H)-one(580 g, in ACN and pyridine was added Ac2O and the reaction mixture washeated to 80° C. for 16 h. After consumption of the starting material(monitored by TLC), the reaction mixture was concentrated in vacuum andcodistilled with toluene (3×250 mL) to afford the title compound as adark brown sticky liquid (595 g, crude, 88%). 1H-NMR (400 MHz; CDCl3): □6.03-6.02 d, 1H), 5.39-5.38 (d, J=5.4 Hz, 1H), 4.7 (t, 1H), 4.0-4.10 (m,2H), 2.23 (s, 3H), 1.54 (s, 3H), 1.46 (s, 3H), 1.44 (s, 3H), 1.37 (s,3H).

Step 3

(3aR,5S,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-yl acetate: To a stirred solution of(3aR,6aR)-5-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyl-3a,6a-dihydrofuro[2,3-d][1,3]dioxol-6-yl acetate (595 g,) in EtOAc (8 volume) wasadded 10% Pd/C (200 g, 50% wet) and the reaction mixture was stirred at40° C. for 12 h under H2 atm (80 psi). After completion, the reactionmixture was filtered through celite, washed with EtOAc (5×300 mL) andconcentrated in vacuum to afford the title compound as a sticky yellowliquid (544 g, 91%). 1H NMR (400 MHz; CDCl3): □ 5.80 (d, J=4.0 Hz, 1H),5.04 (t, J=12.3 Hz, 1H), 4.78-4.81 (m, 1H), 4.58-4.64 (m, 1H), 4.01-4.13(m, 2H), 3.5 (t, J=15.7 Hz, 1H), 2.16 (s, 3H), 1.57 (s, 3H), 1.43 (s,3H), 1.37 (s, 3H), 1.34 (s, 3H).

Step 4

(3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol:To a stirred solution of(3aR,5S,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-yl acetate (544.0 g) in MeOH:H2O(1900 mL:1900 mL) was added Et3N (3.0 eq) and the reaction mixture wasstirred at rt for 3.5 h. After completion, the reaction mixture wasconcentrated in vacuum and codistilled with toluene (3×500 mL) to affordthe title compound as a black solid (510 g, crude). The crude was usedfor next step without purification. 1H NMR (400 MHz; CDCl3): □ 5.78 (d,J=4.0 Hz, 1H), 4.66 (t, J=10.2 Hz, 1H), 4.44-4.50 (m, 1H), 4.2 (m, 1H),3.9 (m, 1H), 3.03-3.09 (m, 1H), 3.70 (t, J=4.5 Hz, 1H), 1.44 (s, 3H),1.42 (s, 3H), 137 (s, 3H).

Steps 5 and 6

(3aR,5R,6S,6aR)-6-azido-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole:

To a stirred solution of(3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxoi-6-ol(510 g) in DCM:pyridine (3.0 eq) was slowly added triflic anhydride inDCM at −20° C. and the reaction mixture was stirred at same temperaturefor 30 min. after completion, the reaction mixture was quenched with icecold 1N HCl (pH ˜6) and the aqueous layer was extracted with DCM (2×1000mL), dried (Na2SO4) and concentrated. This crude residue was dissolvedin DMF and NaN3 (5.0 eq) was added portionwise at 0° C. and stirred atthe same temperature for 3 h. After completion, the reaction mixture waspoured into ice water (500 mL) and extracted with ethyl acetate (2×1000mL). The combined organic layer was again washed with ice cold water(3×500 mL), dried (Na2SO4) and concentrated. The residue was purified byflash column chromatography [normal phase, silica gel (100-200 mesh),gradient 0 to 5% EtOAc in hexane] to afford the title compound as paleyellow gum (150 g, 27%). 1H NMR (400 MHz, CDCl3): □ 5.80 (d, J=3.8 Hz,1H), 4.60-4.63 (m, 1H) 4.35-4.39 (m, 1H), 4.10 (t, J=3.6 Hz, 1H), 3.94(d, J=2.8 Hz, 1H), 3.89-3.93 (m, 2H), 1.58 (s, 3H), 1.55 (s, 3H), 1.45(s, 3H), 1.36 (s, 3H).

Step 7

Synthesis of(3R,4S,5R,6R)-4-azido-6-(hydroxymethyl)tetrahydro-2H-pyran-2,3,5-triol:To a solution of(3aR,5R,6S,6aR)-6-azido-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro [2,3-d][1,3]dioxole (150 g) in DCM (500 mL) 90% TFA inwater was slowly added at −20° C. and stirred at same temperature for 15min. After completion the reaction mixture was concentrated in vacuumand codistilled with toluene (3×500 mL) to afford(3R,4S,5R,6R)-4-azido-6-(hydroxymethyl)tetrahydro-2H-pyran-2,3,5-triol(100 g, 94%) as a yellow solid.

1H-NMR (400 MHz: CDCl3): □ 5.23 (d, J=3.44 Hz, 1H), 4.59 (d, J=7.64 Hz,1H), 4.64 (t, J=15.7 Hz, 2H), 3.87-3.93 (m, 1H), 3.58 (t, J=17.7 Hz,1H), 3.48-3.56 (m, 1H).

Step 1(3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-2,3,5-triol (3)

CuSO4.5H2O (638 mg, 1.64 mmol) and sodium ascorbate (870 mg, 4.39 mmol)were added to a solution of(3R,4S,5R,6R)-4-azido-6-(hydroxymethyl)tetrahydro-2H-pyran-2,3,5-triol(2.0 g, 9.75 mmol) and 1-ethynyl-3-fluorobenzene (2.46 g, 19.51 mmol) inEtOH—H2O (1:1, 20 mL) at room temperature and the reaction mixture washeated to 70° C. for 5 h. After completion, the reaction mixture wascooled to room temperature. The volatiles were evaporated and theaqueous part was extracted with EtOAc (3×30 mL). The organic layer wasdried (Na2SO4) and concentrated and the residue was triturated with Et2Oto afford the title compound as a yellow solid (2.8 g, 90%). ESIMS m/z326 [M+H]+; 1H NMR (400 MHz, DMSO-d6, anomeric mixture, α:β=1:1): d3.37-3.46 (m, 3H), 3.49-3.57 (m, 3H), 3.66 (t, J=6.1 Hz, 1H), 3.86-3.89(m, 1H), 392-3.96 (m, 2H), 4.03 (t, J=6.2 Hz, 1H), 4.25-4.32 (m, 1H),4.53-4.61 (m, 2H), 4.66 (t, J=5.5 Hz, 1H), 4.71 (dd, J=11.0 & 3.1 Hz,1H), 4.84-4.89 (m, 2H), 5.11-5.17 (m, 3H), 5.23 (d, J=5.7 Hz, 1H), 6.71(d, J=4.5 Hz, 1H), 6.93 (d, J=6.1 Hz, 1H), 7.14 (t, J=8.5 Hz, 2H),7.46-7.51 (m, 2H), 7.69 (d, J=10.2 Hz, 2H), 7.73 (d, J=7.8 Hz, 2H), 8.57(s, 1H), 8.61 (s, 1H).

Step 2(2R,3R,4S,5R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-(methylamino)tetrahydro-2H-pyran-3,5-diol(4)

Methyl amine (1.0 M in THF, 10.0 mL) was added a solution of(3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-2,3,5-trial(950 mg, 2.91 mmol) in THF (4 mL), at 0° C. The resulting reactionmixture was stirred at rt for 3h. After completion the volatiles wereevaporate under reduced pressure to the title compound as a greenishsolid (900 mg, crude). ESIMS m/z 347.12 [M+H]+; 1H NMR (400 MHz, CDCl3):□ 2.04 (s, 3H), 2.06 (s, 3H), 2.18 (s, 3H), 2.45 (s, 3H), 2.76-2.80 (m,1H), 4.03-4.17 (m, 3H), 5.44-5.53 (m, 3H), 7.27 (d, J=8.1 Hz, 2H), 7.75(d, J=8.1 Hz, 2H).

Step 1N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylcyclopropanecarboxamide(GTJC-013-03)

Na2CO3 (235 mg, 2.212 mmol) and cyclopropanecarbonyl chloride 2 (94 mg,0.885 mmol) were added to a solution of(2R,3R,4S,5R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-(methylamino)tetrahydro-2H-pyran-3,5-diol1 (150 mg, 0.442 mmol) in methanol (3 mL) at 0° C. The reaction mixturewas stirred at room temperature. After completion the reaction mixturewas quenched with water (5 mL) and extracted with EtOAc (3×25 mL). Thecombined organic layers were washed with brine, dried (Na2SO4), filteredand concentrated under reduced pressure at 45° C. The residue waspurified by flash column chromatography eluting with 4% Methanol in DCMto afford the title compound as a white solid (35 mg, 19%). HRMS: (ESI)[M+H]+ calc. for C19H23FN4O5 406.17, found: 407.36 [M+H]+; LCMS: m/z 407[M+H]+ (ES+) at 89.73% at 3.92 min and 7.08% at 4.14 min.

1H NMR (400 MHz, DMSO-d6, anomeric mixture, α:β=1:9): □ 8.72 (s, 1H),7.69-7.76 (m, 2H), 7.43-7.52 (m, 1H), 7.13-7.17 (m, 1H), 5.54-5.57 (m,1H), 5.34 (d, 0.9H, J1-2=6.4 Hz, α-H-1), 5.33 (d, 0.1H, J1-2=2.7 Hz,p-H-1), 4.96-5.00 (m, 1H), 4.48-4.82 (m, 2H), 3.74-3.92 (m, 2H),3.48-3.53 (m, 2H), 3.13 (s, 3H), 2.08 (m, 1H), 0.75-0.85 (in, 4H),

Step 1N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylthiophene-2-carboxamide(GTJC-013-04)

Na2CO3 (47.04 mg, 0.4437 mmol) and thiophene-2-carbonyl chloride 2(43.19 mg, 0.2958 mmol) were added to a solution of(2R,3R,4S,5R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-(methylamino)tetrahydro-2H-pyran-3,5-diol1 (50 mg, 0.1479 mmol) in methanol (3 mL) at 0° C. The reaction mixturewas stirred at room temperature. After completion the reaction mixturewas quenched with water (5 mL) and extracted with EtOAc (3×25 mL). Thecombined organic layers were washed with brine, dried (Na2SO4), filteredand concentrated under reduced pressure at 45° C. The residue waspurified by flash column chromatography by using 2% Methanol in DCM toafford the title compound as a white solid (15 mg, 23%). HRMS (ESI)[M+H]+ calc. for C20H21FN4O5S: 448.12, found: 449.35 [M+H]+; LCMS: m/z449 [M+H]+;

1H NMR (400 MHz, DMSO-d6, single β isomer): □ 8.66 (s, 1H), 7.83 (d,1H), 7.75-7.82 (m, 2H), 7.63 (d, 1H), 7.47-7.52 (m, 1H), 7.13-7.17 (m,2H), 5.60 (s, 1H), 5.35 (d, J1-2=6.5 Hz, α-H-1), 5.19 (s, 1H), 4.91-4.94(m, 1H), 4.86 (m, 1H), 4.52-4.54 (m, 1H), 3.90 (m, 1H), 3.79 (m, 1H),3.52-3.56 (m, 2H), 3.06 (s, 3H).

Synthesis of G617 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methyl-2-naphthamide(GTJC-013-08)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 85 mg; yield 23%

LCMS: m/z 493 [M+H]+; 1H NMR (400 MHz, DMSO-d6): □ 3.13 (s, 3H),3.52-3.60 (m, 2H), 3.66-3.72 (m, 1H), 3.79-3.83 (m, 1H), 4.50-4.56 (m,1H), 4.72-4.77 (m, 1H), 4.85 (d, J=8.8 Hz, 1H), 5.01 (t, J=5.3 Hz, 1H),5.33 (d, J=6.8 Hz, 1H), 5.55 (d. J=6.4 Hz, 1H), 7.13-7.18 (m, 1H),7.46-7.52 (m, 1H), 7.58-7.74 (m, 5H), 799 (d, J=8.0 Hz, 2H), 8.05 (d,J=7.6 Hz, 1H), 8.25 (s, 1H), 8.71 (s, 1H).

Synthesis of G627 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methyl-3-(trifluoromethyl)benzamide(GTJC-013-09)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 40 mg; yield 13%

HRMS (ESI) [M+H]+ calc. for C23H22F4N4O5 510.15, found: 511.37 [M+H]+

1H NMR (400 MHz, DMSO-d6) (anomeric mixture α:β=1:8): □ 8.73 (s, 1H),7.87-7.93 (m, 3H), 7.67-7.74 (m, 3H), 7.47-7.52 (m, 1H), 7.13-7.18 (m,1H), 5.60 (d, J1-2=6.68 Hz, α-H-1), 5.34 (d, 1H,), 5.01 (d, J1-2=4.7 Hz,β-H-1), 4.43-4.83 (m, 4H), 3.53-3.58 (n, 4H), 3.08 (s, 3H).

Synthesis of G628 (FIG. 4B)3,4-difluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylbenzamide(GTJC-013-10)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 25 mg; yield 9%

α, β isomers are separated by Prep HPLC.

LCMS (β isomer): m/z 479 [M+H]+ (ES+), at 4.65 min (98.44%)

LCMS (α isomer): m/z 479 [M+H]+ (ES+), at 4.79 min (97.98%)

1H-NMR (400 MHz, DMSO-d6, single β isomer) □ 8.73 (s, 1H), 7.72-7.74 (m,1H), 7.62-7.67 (m, 1H), 7.57-7.60 (m, 1H), 7.50-7.54 (m, 2H), 7.42-7.47(m, 1H), 7.12-7.17 (m, 1H), 5.58-5.62 (m, 1H), 5.34 (d, 1H, J=6.68 Hz),4.82-4.99 (m, 2H), 4.78 (d, 1H, J1-2=11.9 Hz, α-H-1), 4.45-4.52 (m, 1H),3.82-3.96 (m, 1H), 3.49-3.60 (m, 3H), 3.05 (s, 3H). 1H-NMR (400 MHz;DMSO-d6, single α isomer) □ 9.00 (s, 1H), 7.72-7.74 (m, 1H), 7.66-7.68(m, 1H), 7.49-7.62 (m, 3H), 7.42 (m, 1H), 7.16-7.21 (m, 1H), 6.21 (bs,1H), 5.17-5.29 (m, 3H), 4.78-4.83 (m, 1H), 4.62-4.64 (m, 1H), 4.45 (d,1H, J1-2=8.12 Hz, β-H-1), 3.38-3.41 (m, 2H), 3.28-3.33 (m, 2H), 3.05 (s,3H).

Synthesis of G622 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methyl-1H-indole-2-carboxamide(GTJC-013-11)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 12 mg; yield 8%

HRMS (ESI) [M+H]+ calc. for C24H24FN5O5 481.18, found: 482.38 [M+H]+;LCMS: m/z 482 (M+H)+ (ES+) 93.64% at 4.78 min; 1H NMR (400 MHz, DMSO-d6,single β isomer): □ 11.58 (s, 1H), 8.77 (s, 1H), 7.71-7.73 (m, 1H),7.62-7.68 (m, 2H), 7.49-7.52 (m, 1H), 7.44-7.47 (m, 1H), 7.19-7.22 (m,1H), 7.13-7.17 (m, 1H), 7.06-7.08 (m, 1H), 7.03 (s, 1H), 5.60 (s, 2H),5.36 (d, 1H, J1-2=6.52 Hz, α-H-1), 4.94 (m, 2H), 4.55 (m, 1H), 3.84-3.91(m, 2H), 3.55-3.64 (m, 2H), 3.10 (s, 3H).

Synthesis of G641 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methyl-2-phenylacetamide(GTJC-013-27)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 30 mg; yield 23%

ESIMS: m/z 347.12 [M+1]+; 1H NMR (400 MHz, DMSO-d6): 3.49-3.61 (m, 4H),3.72 (t, J=6.2 Hz, 2H), 3.99 (dd, 6.6 & 2.9 Hz, 2H), 4.36-4.43 (m, 2H),4.70 (t, J=5.5 Hz, 1H), 4.82 (dd, 10.5 & 2.8 Hz, 2H), 5.19 (d, J=9.7 Hz,2H), 5.31 (d, J=7.2 Hz, 2H), 5.40 (d, J=6.6 Hz, 2H), 7.12-7.17 (m, 2H),7.46-7.51 (m, 2H), 7.66 (dd, J=10.2 & 2.3 Hz, 2H), 7.72 (d, J=7.8 Hz,2H), 8.67 (s, 2H).

Synthesis of G649 (FIG. 4B)2-(3,4-difluorophenyl)-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylacetamide(GTJC-013-37)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 25 mg; yield 17%

HRMS (ESI) [M+H]+ calc. for C23H23F3N4O5 492.16, found: 493.5 [M+H]+

LCMS: m/z 493.5 [M+H]+ (ES+) 88.40% at 4.75 min and 9.81% at 4.88 min,

1H-NMR (400 MHz; DMSO-d6, anomeric mixture, α:β=1:9): □ 8.74 (s, 1H),7.69-7.76 (in, 2H), 7.49-7.53 (m, 1H), 7.31-7.39 (m, 2H), 7.28-7.29 (m,2H), 5.59 (d, 1H, J1-2=9.48 Hz, α-H-1), 4.70-5.36 (m, 5H), 3.76-3.94 (m,4H), 3.45-3.51 (M, 2H), 3.03 (s, 3H).

Synthesis of G651 (FIG. 4B)2-(3,4-difluorophenoxy)-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylacetamide(GTJC-013-38)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 15 mg; yield 11%

HRMS (ESI) [M+H]+ calc. for C23H23F3N4O6 508.16, found: 509.52 [M+H]+

LCMS: m/z 509.5 [M+H]+ (ES+) 91.95% at 4.86 min & 6.98% at 4.96 min,

1H NMR (400 MHz; DMSO-d6, anomeric mixture α:β=1:13): □ 8.77 (s, 2H),7.74-7.76 (m, 2H), 7.69-7.72 (m, 2H), 7.47-7.53 (m, 2H), 7.30-7.37 (m,2H), 7.13-7.18 (m, 2H), 7.02-7.08 (m, 2H), 6.76-6.80 (m, 2H), 5.73 (d,1H, J1-2=6.8 Hz, α-H-1), 5.37-5.41 (m, 2H), 5.29-5.31 (m, 1H), 5.02-5.04(m, 1H), 4.91-4.98 (m, 5H), 4.76-4.79 (m, 2H), 4.70-4.72 (m, 1H),4.37-4.44 (m, 2H), 3.91-3.96 (m, 3H), 3.79-3.82 (m, 1H), 3.50-3.56 (m,2H), 3.00 (s, 2H), 2.88 (s, 3H).

Synthesis of G652 (FIG. 4B)3-(3,4-difluorophenyl)-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylpropanamide(GTJC-013-41)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 11 mg; yield 9%

HRMS (ESI) [M+H]+ calc. for C24H25F3N4O5 506.18, found: 507.52 [M+H]+;LCMS: m/z 507.5 [M+H]+ (ES+) 75.38% at 5.04 min, 7.07% at 5.15 min,7.07%, 15.93% at 5.28 min.

1H NMR (400 MHz; DMSO-d6, mixture of 3 isomers): □ 8.71 (s, 1H),7.51-7.77 (m, 2H), 7.37-7.49 (m, 1H), 7.29-7.35 (m, 2H), 7.14-7.26 (m,2H), 5.48 (d, 1H, J1-2=6.92 Hz, α-H-1), 4.68-5.32 (m, 4H), 4.40-4.50 (m,1H), 3.51-3.99 (m, 4H), 2.54-2.89 (m, 7H).

Synthesis of G658 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-3,4-dimethoxy-N-methylbenzamide(GTJC-013-46-1)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 110 mg; yield 37%

HRMS (ESI) [M+H]+ calc. for C24H27FN4O7 502.19, found: 503.52 [M+H]+;LCMS: m/z 503.5 (M+H)+ (ES+) 95.21% at 4.25 min; 1H NMR (400 MHz;DMSO-d6, single β isomer): □ 8.71 (s, 1H), 7.68-7.75 (m, 2H), 7.46-7.52(m, 1H), 7.12-7.18 (m, 3H), 7.01 (d, J=8.28 Hz, 1H), 5.55 (d, 1H,J1-2=6.36 Hz, α-H-1), 5.32 (d, J=6.64 Hz, 1H), 4.90-4.92 (m, 1H), 4.83(m, 2H), 4.45-4.52 (m, 1H), 3.79-3.85 (m, 7H), 3.61-3.67 (m, 1H),3.32-3.59 (m, 2H), 3.02 (s, 3H).

Synthesis of 655 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-3,4-dihydroxy-N-methylbenzamide(GTJC-013-46)

Synthesized following the standard procedure used for GTJC-013-23

Appearance: white solid; synthesized: 18 mg; yield 24%

HRMS (ESI) [M+H]+ calc. for C22H23FN4O7 474.16, found: 475.50 [M+H]+

LCMS: m/z 475.5 [M+H]+ (ES+) 98.93% at 3.86 min.

1H NMR (400 MHz; DMSO-d6, single β isomer): □ 9.11 (bs, 2H), 8.71 (s,1H), 7.74 (d, 1H, J=7.68 Hz,), 7.69 (d, J=10.32 Hz, 1H), 7.46-7.52 (m,1H), 7.12-7.17 (m, 1H), 6.94 (s, 2H), 6.73-6.75 (m, 1H), 5.51 (d, 1H,J1-2=6.01 Hz, α-H-1), 5.29 (bs, 1H), 4.45-4.85 (m, 4H), 3.86 (bs, 1H),3.57 (m, 3H), 2.98 (s, 3H).

Synthesis of G642 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methyl-3-(trifluoromethoxy)benzamide(GTJC-013-45)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 50 mg; yield 33%

HRMS (ESI) [M+H]+ calc. for C23H22F4N4O6 526.15, found: 527.47 [M+H]+

1H NMR (400 MHz, DMSO-d6, anometic mixture): d 8.82 (s, 1H), 7.83-7.93(m, 3H), 7.69-7.77 (m, 3H), 7.49-7.52 (m, 1H), 7.13-7.18 (m, 1H), 5.60(d, 1H, J1-2=6.68 Hz, α-H-1), 5.34 (d, J=6.6 Hz, 1H), 5.14 (d, 1H,J1-2=4.0 Hz, β-H-1), 4.47-4.61 (m, 4H), 3.39-3.60 (m, 4H), 3.08 (s, 3H).

Synthesis of G650 (FIG. 4B) Synthesis of2,3,4,5,6-pentafluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylbenzamide(GTJC-013-47)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 30 mg; yield 19%

HRMS (ESI) [M+H]+ calc. for: C22H18F6N4O5 532.12, found: 533.48 [M+H]+;LCMS: m/z 533.4 [M+H]+ (ES+) 82.08% at 504 min & 14.98% at 5.15 min.

1H NMR (400 MHz, DMSO-d6, anomeric mixture, α:β=1:6): □ 8.71 (s, 1H),7.55-7.74 (m, 2H), 7.47-7.52 (m, 1H), 7.13-7.21 (m, 1H), 5.37 (d, 1H,J1-2=6.6 Hz, α-H-1), 4.37-5.62 (m, 5H), 3.32-3.69 (m, 4H), 3.07 (s, 3H).

Synthesis of G629 (FIG. 4B)N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1-methoxy-N-methyl-2-naphthamide(GTJC-013-22)

Synthesized following the standard procedure used for GTJC-013-03 orGTJC-013-04

Appearance: white solid; synthesized: 110 mg; yield 35%

ESIMS: m/z 347.12 [M+H]+; 1H NMR (400 MHz, DMSO-d6): □ 3.49-3.61 (m,4H), 3.72 (t, J=6.2 Hz, 2H), 3.99 (dd, 6.6 & 2.9 Hz, 2H), 4.36-4.43 (m,2H), 4.70 (t, J=5.5 Hz, 1H), 4.82 (dd, 10.5, 2.8 Hz, 2H), 5.19 (d, J=9.7Hz, 2H), 5.31 (d, J=7.2 Hz, 2H), 5.40 (d, J=6.6 Hz, 2H), 7.12-7.17 (m,2H), 7.46-7.51 (m, 2H), 7.66 (dd, J=10.2 & 2.3 Hz, 2H), 7.72 (d, J=7.8Hz, 2H), 8.67 (s, 2H).

N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1-hydroxy-N-methyl-2-naphthamide(GTJC-013-23)

To a solution ofN-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1-methoxy-N-methyl-2-naphthamide(80 mg, 0.1532 mmol) in DCM (4 mL) added BBr3 (115.4 mg, 0.4597 mmol) at0° C. The resulting mixture was stirred at room temperature for 3 h.After completion, the reaction mixture was quenched with saturatedNaHCO₃ solution (6 mL) to adjust pH˜8 and the aqueous layer wasextracted with DCM (3×10 mL). The combined organic layer was dried(Na2SO4) and concentrated in vacuo. The residue was purified by Flashchromatography eluting with 3% Methanol in DCM to afford the titlecompound as white solid (13 mg, 17%). ESIMS: m/z 347.12 [M+1]+; 1H NMR(400 MHz, DMSO-d6): □ 3.49-3.61 (m, 4H), 3.72 (t, J=6.2 Hz, 2H), 3.99(dd, 6.6, 2.9 Hz, 2H), 4.36-4.43 (m, 2H), 4.70 (t, J=5.5 Hz, 1H), 4.82(dd, 10.5 & 2.8 Hz, 2H), 5.19 (d, J=9.7 Hz, 2H), 5.31 (d, J=7.2 Hz, 2H),5.40 (d, J=6.6 Hz, 2H), 7.12-7.17 (m, 2H), 7.46-7.51 (m, 2H), 7.66 (dd,J=10.2 & 2.3 Hz, 2H), 7.72 (d, J=7.8 Hz, 2H), 8.67 (s, 2H).

N1,N4-bis((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N1,N4-dimethylterephthalamide(GTJC-013-12): To a solution of(2R,3R,4S,5R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-(methylamino)tetrahydro-2H-pyran-3,5-diol(140 mg, 0.4142 mmol) in methanol (3 mL) was added Na2CO3 (220 mg,2.0710 mmol) and terephthaloyl dichloride (172 mg, 0.8284 mmol) at 0° C.The reaction mixture was stirred at room temperature. After completionthe reaction mixture was quenched with water (5 mL) and extracted withEtOAc (3×25 mL). The combined organic layers were washed with brine anddried (Na2SO4), filtered and concentrated under reduced pressure at 45°C. The residue was purified by Prep HPLC to afford the title compound (3mg) as white solid. HRMS (ESI) [M+H]+ calc. for C38H40F2N8010 806.28,found: 807.71 [M+H]+; LCMS: m/z 807.7 (M+H)+ (ES+) 98.10% at 4.49 min.1H NMR (400 MHz; DMSO-d6, βisomer): □ 8.73 (s, 2H), 7.65-7.74 (m, 8H),7.47-7.52 (m, 2H), 7.13-7.17 (m, 2H), 5.64 (d, 2H, J1-2=6.48 Hz, α-H-1),5.34-5.36 (m, 2H), 4.76-4.81 (m, 4H), 4.49-4.53 (m, 2H), 3.61-3.73 (m,2H), 3.54-3.58 (m, 6H), 3.08 (s, 6H).

Step-1(2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-azido-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate

To a solution of(2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-bromo-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate (1.92 g, 3.74 mmol) in DMF (20 mL), NaN3 (1.21 g, 18.7 mmol)was added at room temperature. The reaction mixture was heated to 80 0Cfor 3 h. After completion, the reaction mixture was cooled to roomtemperature and quenched with cold water (20 mL). The aqueous layer wasextracted with EtOAc (2×20 mL), dried (Na2SO4) and concentrated invacuo. The crude residue was purified by flash column chromatography[normal phase, silica gel (100-200 mesh), gradient 0 to 50% EtOAc inhexane] to afford the title compound as a white solid (670 mg, 38%).ESIMS m/z 477 [M+H]+; 1H NMR (400 MHz, CDCl3): δ 1.95 (s, 3H), 2.07 (s,6H), 4.17-4.24 (m, 3H), 4.81 (d, J=8.5 Hz, 1H), 5.17 (dd, J=11.3 & 3.1Hz, 1H), 5.59 (d, J=2.9 Hz, 1H), 5.62-5.70 (m, 1H), 7.02-7.06 (m, 1H),7.35-7.41 (m, 1H), 7.52 (t, J=7.7 Hz, 2H), 7.80 (s, 1H).

Step-2(3R,4S,5R,6R)-2-(2-naphthamido)-6-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate

To a solution of(2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-azido-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyl diacetate (100 mg, 0.21 mmol) in THF (5 mL),Pd—C (20 mg, 10%, dry) was added and the reaction mixture was stirredunder H2 (1 atm) at room temperature for 2 h. After completion, pyridine(0.05 mL, 0.63 mmol) was added to the reaction mixture, cooled to 0° C.and 2-naphthoyl chloride (80 mg, 0.42 mmol) was slowly added and stirredat room temperature for 2 h. After completion, the reaction mixture wasfiltered, washed with EtOAc (3×10 mL). The combined organic layers werewashed with water (10 mL), dried (Na2SO4), and concentrated in vacuo toafford the title compound as a white sticky solid (158 mg, crude).ESIMS: m/z 605 [M+H]+.

Step-3N-((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-2-naphthamide(GTJC-013-15)

To a solution of(3R,4S,5R,6R)-2-(2-naphthamido)-6-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate (4, 158 mg, 0.26 mmol) MeOH (5 mL), NaOMe (0.26 mL, 1M, 0.26mmol) was added at 0° C. The reaction mixture was stirred at 0° C. for 2h. After completion, the reaction mixture was acidified with Amberlyst15 (pH ˜6) and filtered. Washed with MeOH (3×10 mL) and concentrated invacuo. The residue was purified by flash column chromatography [normalphase, silica gel (100-200 mesh), gradient 0 to 10% MeOH in DCM] toafford the title compound as a white solid (60 mg, 48%). LCMS: m/z 479(M+H)+; (ES+) 70.96% at 4.80 min and 23.57% at 4.87 min, 1H NMR (400MHz; DMSO-d6): □ 3.53 (t, J=5.9 Hz, 2H), 3.83 (t, J=6.1 Hz, 1H),3.97-4.02 (m, 2H), 4.39-4.44 (m, 1H), 4.70-4.74 (m, 1H), 4.93 (dd,J=10.8, 2.8 Hz, 1H), 5.28 (d, J=6.7 Hz, 1H), 5.36 (d, J=6.7 Hz, 1H),7.16 (td, J=8.8, 2.5 Hz, 1H), 7.46-7.52 (m, 1H), 7.59-7.66 (m, 2H),7.69-7.76 (m, 2H), 7.96-8.07 (m, 4H), 8.50, 8.60 (each singlet, 1H),9.27 (d, J=8.9 Hz, 1H).

Step-1(3R,4S,5R,6R)-2-(benzylamino)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,5-diol

Benzylamine (87.1 mg, 0.8136 mmol) was added to a solution of(3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetra hydro-2H-pyran-2,3,5-triol (250 mg, 0.7396 mmol) in THF (2 mL) andthe mixture was stirred at room temperature for 2 h. After completion,the reaction mixture was concentrated and the residue was triturated byEt2O to afford the title compound as a light yellow solid (150 mg). Thecrude material was used in next step. ESIMS: m/z 353 [M+H]+.

Step 2N-benzyl-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-2-naphthamide

Na2CO3 (115.04 mg, 1.0869 mmol) and 2-naphthoyl chloride (190.99 mg,0.7246 mmol) were added to a solution of(3R,4S,5R,6R)-2-(benzylamino)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,5-diol(150 mg, 0.3623 mmol) in methanol (3 mL) at 0° C. The reaction mixturewas stirred at room temperature. After completion, the reaction mixturewas quenched with water (5 mL) and extracted with EtOAc (3×25 mL). Thecombined organic layers were washed with brine, dried (Na2SO4), filteredand concentrated under reduced pressure at 45° C. The residue waspurified by prep HPLC to afford anomeric mixture the title compound aswhite solid (3 mg). HRMS (ESI) [M+H]+ calc. for C32H29FN4O5 568.21,found: 569.53 [M+H]+; LCMS: m/z 569.5 [M+H]+(ES+) 96.83% at 5.59 min.

1H NMR (400 MHz; DMSO-d6, single β isomer): 7 8.70 (s, 1H), 8.24 (s,1H), 8.07-8.08 (m, 1H), 7.98-8.01 (m, 2H), 7.51-7.71 (m, 7H), 7.46-7.51(m, 1H), 7.31-7.35 (m, 2H), 7.20-7.24 (m, 1H), 7.12-7.17 (m, 1H), 5.66(bs, 1H), 5.32 (bs, 1H), 5.06 (d, 1H, J1-2=7.72 Hz, α-H-1), 4.86-4.89(m, 1H), 4.77-4.79 (m, 1H), 4.65-4.69 (m, 1H), 4.54-4.59 (m, 1H), 3.83(s, 1H), 3.62-3.70 (m, 2H), 3.56-3.58 (m, 1H).

Synthesis of G639N-benzyl-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)benzamide(GTJC-013-20)

Synthesized following the standard procedure used for GTJC-013-24

Appearance: white solid; synthesized: 1 mg

ESIMS: 949 [M+H]+; LCMS: m/z 697 (M+H)+ (ES+) 96.37% at 4.51 min,

1H NMR (400 MHz, DMSO-d6): □ 3.49-3.61 (m, 4H), 3.72 (t, J=6.2 Hz, 2H),3.99 (dd, 6.6 & 2.9 Hz, 2H), 4.36-4.43 (m, 2H), 4.70 (t, J=5.5 Hz, 1H),4.82 (dd, 10.5 & 2.8 Hz, 2H), 5.19 (d, J=9.7 Hz, 2H), 5.31 (d, J=7.2 Hz,2H), 5.40 (d, J=6.6 Hz, 2H), 7.12-7.17 (m, 2H), 7.46-7.51 (m, 2H), 7.66(dd, J=10.2 & 2.3 Hz, 2H), 7.72 (d, J=7.8 Hz, 2H), 8.67 (s, 2H).

Step 1(3R,4S,5R,6R)-2-(ethylamino)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,5-diol(2)

A solution of(3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetra hydro-2H-pyran-2,3,5-triol (3, 150 mg, 1.53 mmol) in ethylamine(1.0 M in THF, 2 mL) was stirred at room temperature for 2 h. Aftercompletion, the reaction mixture was concentrated in vacuo. Theresulting crude residue was triturated by Et2O to afford the titlecompound as a light yellow solid (100 mg, crude). The material was takenfor next step without further purification. HRMS (ESI) [M+H]+ calc. forC16H21FN4O4 352.15, found: 353.33 [M+H]+; ESIMS: m/z 353 [M+H]+.

Step-2N-ethyl-3,4-difluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)benzamide(GTJC-013-42)

To a solution of(3R,4S,5R,6R)-2-(ethylamino)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,5-diol(100 mg, 0.2840 mmol) in methanol (3 mL) was added Na2CO3 (90.04 mg,0.8522 mmol) and 3,4-difluorobenzoyl chloride (99.99 mg, 0.5681 mmol) at0° C. The reaction mixture was stirred at room temperature. After 3 h,the reaction mixture was quenched with water (5 mL) and extracted withEtOAc (3×25 mL). The combined organic layers were washed with brine anddried (Na2SO4). The solvent was removed under reduced pressure at 45° C.and the residue was purified by flash column chromatography by using4.5% Methanol in DCM to afford the title compound as white solid (15 mg,11%).

HRMS (ESI) [M+H]+ calc. for C23H23F3N4O5 492.16, found: 493.47 [M+H]+

LCMS: m/z 493.4 [M+H]+ (ES+), at 5.28 min (93.45%) & 5.39 min (6.26%).

1H NMR (400 MHz; DMSO-d6, anomeric mixture α:β=1:15): □ 8.75 (s, 1H),7.71-7.76 (m, 1H), 7.65-7.69 (m, 1H), 7.48-7.59 (m, 3H), 7.41-7.47 (m,1H), 7.12-7.17 (m, 1H), 5.50 (d, 1H, J1-2=6.92 Hz, α-H-1), 5.34 (d,J=6.48 Hz, 1H), 4.62-4.79 (m, 3H), 4.42-4.49 (m, 1H), 3.82 (bs, 1H),3.66-3.69 (m, 1H), 3.32-3.54 (m, 4H), 1.25 (m, 3H).

Step 1(2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-((2-methoxyethyl)amino)tetrahydro-2H-pyran-3,5-diol(3)

2-methoxyethan-1-amine 2 (93.25 mg, 1.226 mmol) was added to a solutionof(3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(hydroxymethyl)tetra hydro-2H-pyran-2,3,5-triol (1, 200 mg, 0.613 mmol) in THF (5 ml)and the mixture was stirred at room temperature for 2 h. Aftercompletion, the reaction mixture was concentrated in vacuo and theresidue was triturated by Et2O to afford the title compound as a lightyellow solid (170 mg, crude. The material was taken for next step. HRMS(ESI) [M+H]+ calc. for C17H23FN4O5 382.17, found: 383.17 [M+H]+; ESIMS:m/z 383 [M+H]+.

Step 23,4-difluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-(2-methoxyethyl)benzamide(GTJC-013-43-1)

To a solution of(2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-((2-methoxyethyl)amino)tetrahydro-2H-pyran-3,5-diol(150 mg, 0.3926 mmol) in methanol (3 mL) was added Na2CO3 (124.8 mg,1.1780 mmol) and 3,4-difluorobenzoyl chloride (138.02 mg, 0.7853 mmol)at 0° C. The reaction mixture was stirred at room temperature. After 3h, the reaction mixture was quenched with water (5 mL) extracted withEtOAc (3×25 mL), The combined organic phase was washed with brine anddried (Na2SO4). The solvent was removed under reduced pressure at 45° C.and the residue was purified by flash column chromatography by using 2%Methanol in DCM to afford the title compound as a white solid (110 mg,54%). HRMS (ESI) [M+H]+ calc. for C24H25F3N4O6 522.17, found: 523.58[M+H]+

1H NMR (400 MHz, DMSO-d6, with D20, β isomer): □ 8.76 (s, 1H), 7.71-7.75(m, 1H), 7.68-7.71 (m, 1H), 7.56-7.68 (m, 3H), 7.47-7.51 (m, 1H),7.12-7.17 (m, 1H), 5.47 (d, 1H, J1-2=6.44 Hz, α-H-1), 5.36 (d, J=6.56Hz, 1H), 4.83-4.85 (m, 1H), 4.76-4.78 (m, 2H), 4.43-4.50 (m, 1H),3.83-3.88 (m, 2H), 3.48-3.56 (m, 6H), 3.29 (s, 3H).

Step 33,4-difluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-(2-hydroxyethyl)benzamide(GTJC-013-43)

To a solution of3,4-difluoro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-(2-methoxyethyl)benzamide(80 mg, 0.1532 mmol) in DCM (4 mL) BBr3 (115.4 mg, 0.4597 mmol) wasadded at 0° C. The reaction mixture was stirred at room temperature for3 h. After completion, the reaction mixture was quenched with saturatedNaHCO₃ solution (6 mL) to adjust pH˜8 and extracted with DCM (3×25 mL).The combined organic phase was washed with brine and dried (Na2SO4)filtered and concentrated under reduced pressure at 45° C. The residuewas purified by prep HPLC to afford the title compound as a white solid(6 mg, 8%).

HRMS (ESI) [M+H]+ calc. for C23H23F3N4O6 508.16, found: 509.5 [M+H]+

LCMS: m/z 509.5 (M+H)+ (ES+) 59.89% at 4.57 min, 29.86% at 4.66 min,9.66% at 4.76 min.

1H NMR (400 MHz; DMSO-d6, mixture of 3 isomers): □ 8.69 (s, 1H),7.45-7.77 (m, 6H), 7.13-7.18 (m, 1H), 5.56 (d, 1H, J1-2=6.16 Hz, α-H-1),5.37 (d, J=6.4 Hz, 1H), 4.41-5.04 (m, 5H), 3.48-4.28 (m, 8H).

Step 1(2R,3R,4S,5R)-2-(acetoxymethyl)-6-amino-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate hydrochloride (3)

10% Pd—C (50 mg) and conc. HCl (two drops) were added solution of(2R,3R,4S,5R)-2-(acetoxymethyl)-6-azido-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate (115 mg, 0.2415 mol) in Methanol (3 mL). The mixture wasstirred under hydrogen atmosphere (balloon pressure) at room temperaturefor 2 h. After completion, the reaction mixture was filtered throughcelite, washed with methanol (10 mL). The combined filtrate wasconcentrated in vacuo to afford the title compound as an off white solid(93 mg, 86%). The residue was used in next step without furtherpurification. 1H NMR (400 MHz, CDCl3): □ 2.04 (s, 3H), 2.06 (s, 3H),2.18 (s, 3H), 2.45 (s, 3H), 2.76-2.80 (m, 1H), 4.03-4.17 (m, 3H),5.44-5.53 (m, 3H), 7.27 (d, J=8.1 Hz, 2H), 7.75 (d, J=8.1 Hz, 2H).

Step 2(2R,3R,4S,5R,6R)-2-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(phenylsulfonamido)tetrahydro-2H-pyran-3,5-diyldiacetate (4)

To a solution of(2R,3R,4S,5R)-2-(acetoxymethyl)-6-amino-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate hydrochloride (3, 93 mg 0.1913 mmol) in DCM (5 mL) pyridine(46 mg, 0.5740 mmol) and benzene sulfonyl chloride chloride (50.69 mg,0.2870 mmol) were added at 0° C. The reaction mixture was stirred atroom temperature for 3 h. After completion, the reaction mixture wasquenched with water (3 mL) and extracted with DCM (3×15 mL). Thecombined organic layers were dried (Na2SO4) and concentrated in vacuo toafford the title compound as an off white semi solid (130 mg, crude).HRMS (ESI) [M+H]+ calc. for C26H27FN4O9S 590.15, found: 591.32 [M+H]+;1H NMR (400 MHz, DMSO-d6): □ 3.49-3.61 (m, 4H), 3.72 (t, J=6.2 Hz, 2H),3.99 (dd, 6.6 & 2.9 Hz, 2H), 4.36-4.43 (m, 2H), 4.70 (t, J=5.5 Hz, 1H),4.82 (dd, 10.5 & 2.8 Hz, 2H), 5.19 (d, J=9.7 Hz, 2H), 5.31 (d, J=7.2 Hz,2H), 5.40 (d, J=6.6 Hz, 2H), 7.12-7.17 (m, 2H), 7.46-7.51 (m, 2H), 7.66(dd, J=10.2 & 2.3 Hz, 2H), 7.72 (d, J=7.8 Hz, 2H), 8.67 (s, 2H).

Step 3N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)benzenesulfonamide(GTJC-026)

To a solution of(2R,3R,4S,5R,6R)-2-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-6-(phenylsulfonamido)tetrahydro-2H-pyran-3,5-diyldiacetate (120 mg, 0.203 mmol) in MeOH (5 mL), NaOMe (0.46 mL, 1M, 0.46mmol) was slowly added at 0° C. The reaction mixture was stirred at roomtemperature for 2 h. After completion, the reaction mixture wasacidified with Amberlyst 15 resin (pH ˜5) filtered, washed with MeCOH(3×5 mL) and concentrated in vacuo. The residue was purified by prepHPLC to afford anomeric mixture of the title compound (GTJC-026-P1, 3mg, mixture of 4 isomers, GTJC-026-P2, 1 mg, anomeric mixture, α:β=1:12)as a white solids.

HRMS (ESI) [M+H]+ calc. for C20H21FN4O6S 464.12, found: 465.42 [M+H]+

LCMS: (GTJC-026-P1) m/z 465.4 [M+H]+ (ES+) 58.97% at 4.33 min, 27.97% at4.38 min, 6.57% at 4.55 min & 2.61% 4.74 min.

1H NMR (GTJC-026-P1) (400 MHz; DMSO-d6, mixture of 4 isomers): 8.60 (s,0.5H), 8.58 (s, 0.5H), 7.87-7.90 (m, 2H), 7.62-7.74 (m, 2H), 7.46-7.61(m, 4H), 7.12-7.16 (m, 1H), 5.38 (d, 1H, J1-2=7.12 Hz, α-H-1), 3.63-5.39(m, 7H), 3.10-3.13 (m, 2H), 2.49-2.58 (m, 1H).

LCMS: (GTJC-026-P2) m/z 465.38 [M+H]+, (ES+) 89.0% at 4.33 min and 7.34%at 4.38 min.

1H NMR (GTJC-026-P2) (400 MHz; DMSO-d6, anomeric mixture, α:β=1:12): □8.76 (bs, 1H), 8.48 (s, 1H), 7.89 (d, J=7.32, 2H), 7.68-7.72 (m, 2H),7.46-7.62 (m, 4H), 7.12-7.17 (m, 1H), 5.40 (d, 1H, J1-2=7.12 Hz, α-H-1),5.26 (d, J=5.7, 1H), 4.83-4.86 (m, 1H), 4.69 (d, J=8.56, 1H), 4.40-4.52(m, 1H), 3.89-3.90 (in, 1H), 3.62-3.65 (m, 1H), 3.09-3.14 (m, 1H), 2.50(s, 1H).

Synthesis of G630N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)benzenesulfonamide(GTJC-055)

Synthesized following the standard procedure used for GTJC-026

Appearance: White solid

Synthesized: (GTJC-055-P1, 3 isomers, 15 mg) and (GTJC-055-P2, 4isomers, 8 mg)

HRMS (ESI) [M+H]+ calc. for C20H19F3N4O6S 500.10, found: 501.20 [M+H]+

LCMS (GTJC-055-P1): m/z 501 [M+H]+ (ES+), 78.32% at 4.66 min, 7.41% at4.72 min & 12.44% 4.83 min.

1H NMR (GTJC-055-P1) (400 MHz; DMSO-d6): □ 8.55 (s, 1H), 7.81-7.91 (m,1H), 7.60-7.74 (m, 4H), 7.46-7.51 (m, 1H), 7.12-7.17 (m, 1H), 5.43 (d,1H, J1-2=7.12 Hz, α-H-1), 5.26 (d, 1H, J=5.88 Hz), 4.83-4.86 (m, 1H),4.67-4.69 (m, 1H), 4.54-4.57 (m, 1H), 4.01-4.04 (m, 1H), 3.89 (bs, 1H),3.66-3.69 (m, 1H), 3.32-3.37 (m, 1H), 3.16-3.19 (m, 1H).

LCMS (GTJC-055-P2,): m/z 501 [M+H]+ (ES+), 62.43% at 4.66 min, 24.51% at4.72 min, 5.63% 4.85 min & 3.81% 5.04 min.

1H NMR (GTJC-055-P2) (400 MHz, DMSO-d6) 8.50 (s, 1H), 7.83-7.94 (m, 1H),7.60-7.74 (m, 5H), 7.46-7.51 (m, 1H), 7.17-7.27 (m, 1H), 5.43 (d, 1H,J1-2=7.2 Hz, α-H-1), 5.24-5.27 (m, 1H), 4.83-4.86 (m, 1H), 4.68 (d,J=8.5 Hz, 1H), 4.46-4.57 (n, 1H), 4.02-4.04 (m, 1H), 3.89 (m, 1H),3.66-3.67 (n, 1H), 3.31-3.35 (m, 1H), 3.16-3.19 (m, 1H).

Step 1

N-(3-mercaptophenyl)acetamide (2): To a solution of 3-aminobenzenethiol(2 g, 16.0 mmol) in EtOAc (50 mL), Ac2O (1.66 mL, 17.6 mmol) was slowlyadded at 0° C. and the reaction mixture was stirred at room temperaturefor 2 h. After completion, the reaction mixture was quenched with water(40 mL). After separating the organic layer, the aq layer was extractedwith EtOAc (2×10 mL). The combined organic layers were washed withbrine, dried (Na2SO4) and concentrated in vacuo to afford the titlecompound as light brown sticky solid (2.23 g, 83%). ESIMS: m/z 166[M+H]+; 1H NMR (400 MHz, DMSO-d6): □ 2.02 (s, 3H), 5.39 (bs, 1H), 6.94(d, J=7.6 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.60(s, 1H), 9.90 (s, 1H).

Step 2(3R,4S,5R,6R)-2-((3-acetamidophenyl)thio)-6-(acetoxymethyl)-4-azidotetrahydro-2H-pyran-3,5-diyldiacetate (4)

A solution of N-(3-mercaptophenyl)acetamide (2, 161 mg, 0.96 mmol) and(3R,4S,5R,6R)-6-(acetoxymethyl)-4-azidotetrahydro-2H-pyran-2,3,5-triyltriacetate (180 mg, 0.48 mmol) in DCM (20 mL), BF3.Et2O (304 mg, 0.96mmol) was slowly added at 0° C. and the reaction mixture was heated at55° C. for 16 h. After completion, the reaction mixture was quenchedwith water (10 mL) and extracted with DCM (3×10 mL). The combinedorganic layers were washed with brine, dried (Na2SO4) and concentratedin vacuo. The crude residue was purified by flash column chromatography[normal phase, silica gel (100-200 mesh), gradient 0% to 70% EtOAc inhexane] to afford the title compound as a off white solid (177 mg, 77%).The crude residue was used for the next step without furtherpurification. ESIMS: m/z 481 [M+H]+

Step 3(3R,4S,5R,6R)-2-((3-acetamidophenyl)thio)-6-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate (6)

To a solution of(3R,4S,5R,6R)-2-((3-acetamidophenyl)thio)-6-(acetoxymethyl)-4-azidotetrahydro-2H-pyran-3,5-diyldiacetate (230 mg 0.48 mmol) and 1-ethynyl-3-fluorobenzene (121 mg, 0.96mmol) in EtOH (5 mL) and water (5 mL), sodium ascorbate (43 mg, 0.21mmol) and CuSO4.5H2O (32 mg, 0.07 mmol) were added at room temperature.The reaction mixture was heated to 70° C. for 2 h. After completion, thereaction mixture was diluted with EtOAc (10 mL) and filtered through apad of celite, washed with EtOAc (3×10 mL), and concentrated in vacuo toafford the title compound as an off white solid (243 mg, 84%). The cruderesidue was used for the next step without further purification. ESIMS:m/z 601 [M+H]+,

Step 4 Synthesis ofN-(3-(((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)thio)phenyl)acetamide(GTJC-023)

To a solution of(3R,4S,5R,6R)-2-((3-acetamidophenyl)thio)-6-(acetoxymethyl)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)tetrahydro-2H-pyran-3,5-diyldiacetate (180 mg, 0.3 mmol) in MeOH (5 mL), NaOMe (0.3 mL, 1 M, 0.3mmol) was slowly added at 0° C. and stirred at rt for 3 h. Aftercompletion, the reaction mixture was acidified with Amberlyst 15 resin(pH ˜5), filtered, washed with MeOH (3×10 mL) and concentrated in vacuo.The crude residue was purified by flash column chromatography [normalphase, silica gel (100-200 mesh), gradient 0% to 10% MeOH in DCM]. Thewhite solid obtained was triturated with Et2O to afford the titlecompound as a white solid (68 mg, 48%).

LCMS: m/z 475 (M+H)+ (ES+), 75.88% at 4.40 min and 22.76% at 4.63 min.

1H NMR (400 MHz; DMSO-d6): □ 2.04 (S, 3H), 3.35-4.05 (overlappingsignals, m, 4H), 4.12-5.67 (overlapping signals, m, 6H), 7.23-7.31 (m,3H), 7.36-7.52 (m, 2H), 7.67-7.76 (m, 2H), 7.80 (s, 1H), 8.69, 8.88(each singlet, 1H), 9,96, 10.01 (each singlet, 1H).

Step 5N-(3-(((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)sulfonyl)phenyl)acetamide(GTJC-029)

To a solution ofN-(3-(((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)thio)phenyl)acetamide (30 mg, 0.063 mmol) inDCM (4 mL), m-CPBA (15 mg, 0.063 mmol) was added at 0° C. and stirred atsame temperature for 2 h. After completion, the reaction mixture wasquenched with aq. NaOH (10 mL, 2M) and the aq. layer was extracted withDCM (3×10 mL). The combined organic layers were washed with brine, dried(Na2SO4) and concentrated in vacuo. The residue was triturated with Et2Oto afford the title compound as a white solid (16 mg, 50%).

LCMS: m/z 475 [M+H]+ (ES+), 22.84% at 4.18 min, 64.79% at 4.29 min,8.33% at 4.41 min, 2.40% at 4.91 min.

1H NMR (400 MHz, DMSO-d6): □ 2.07, 2.08 (each singlet, 3H), 3.26-3.31(m, 1H), 3.40-4.74 (overlapping signals, m, 6H), 4.96 (dd, J=10.6 & 2.3Hz, 1H), 5.22-5.98 (overlapping signals, m, 2H), 7.13-7.17 (m, 1H),7.46-7.62 (m, 3H), 7.67-7.78 (m, 2H), 7.83-7.88 (m, 1H), 8.20, 8.21,8.27 (each singlet, 1H), 8.67, 8.68, 8.77 (each singlet, 1H), 10.29,10.30, 10.34 (each singlet, 1H).

Steps 1 and 2N-((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylpropiolamide(GTJC-013-62)

To a solution of(2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-2-(hydroxymethyl)-6-(methylamino)tetrahydro-2H-pyran-3,5-diol(300 mg, 0.8875 mmol) in MeOH (10 mL), Na2CO3 (940 mg, 8.8757 mmol) wasadded and the reaction mixture was cooled to 0° C. propioloyl chloride(156 mg, 1.7751 mmol) was slowly added at 0° C. and the reaction Mixturewas stirred at room temperature for 2 h. After completion, the reactionmixture was concentrated in vacuo. The crude residue was purified byflash column chromatography [normal phase, silica gel (100-200 mesh),gradient 0 to 10% MeOH in DCM] to give mixture ofN-((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylpropiolamide(GTJC-013-62, single β isomer, 2 mg) as a white solid,(Z)-3-chloro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylacrylamide(GTJC-013-63-P1, single β isomer, 2 mg) as a white solid and(E)-3-chloro-N-((2R,3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylacrylamide(GTJC-013-63-P2, single β isomer, 1 mg) as a white solid.

HRMS (ESI) [M+H]+ calc. for C18H19FN4O5 390.13, found: 391.16 [M+H]+

LCMS (GTJC-013-62): m/z 391 [M+H]+ (ES+) 95.11% at 3.88 min.

1H-NMR (400 MHz; DMSO-d6, (GTJC-013-62, single β isomer)): 8.74 (s, 1H),7.73-7.75 (m, 1H), 7.68-7.70 (m, 1H), 7.47-7.52 (m, 1H), 7.13-7.17 (m,1H), 5.66 (d, 1H, J1-2=6.12 Hz, α-H-1), 3.80-5.51 (m, 7H), 3.51-3.53 (m,2H), 3.15-3.17 (m, 2H), 2.89 (s, 2H).

HRMS (ESI) [M+H]+ calc. for C18H20CIFN4O5 426.11, found: 427.11 [M+H]+LCMS (GTJC-013-63-P1): m/z 427.1 [M+H]+ (ES+) 93.52% at 3.96 min.

1H NMR (400 MHz, DMSO-d6, (GTJC-013-63-P1, single β isomer)): 8.73 (s,1H), 7.53-7.7 5 (m, 1H), 7.68-7.70 (m, 1H), 7.47-7.52 (m, 1H), 7.13-7.17(m, 1H), 6.74-6.76 (m, 1H), 6.65-6.67 (m, 1H), 5.60 (d, 1H, J1-2=8.6 Hz,α-H-1), 5.30-5.37 (m, 1H), 4.91-4.95 (m, 2H), 4.74 (m, 1H), 4.37 (m,1H), 3.92 (m, 1H), 3.81 (t, J=5.96 Hz, 1H), 3.51 (m, 2H), 2.90 (s, 3H).

HRMS (ESI) [M+H]+ calc. for C18H20CIFN4O5 426.11, found: 427.14 [M+H]+

LCMS (GTJC-013-63-P2): m/z 427.1 [M+H]+ (ES+), 90.74% at 4.22 min.

1H NMR (400 MHz; DMSO-d6) (GTJC-013-63-P2, single β isomer): 8.74 (s,1H), 7.73-7.75 (m, 1H), 7.68-7.71 (m, 1H), 7.47-7.52 (m, 1H), 7.31-7.34(m, 1H), 7.13-7.17 (m, 1H), 7.09 (m, 1H), 5.59 (m, 1H), 5.38 (m, 1H),5.22 (d, 1H, J1-2=8.84 Hz, α-H-1), 4.98 (m, 1H), 4.76 (m, 1H), 4.39 (m,1H), 3.92-3.93 (m, 2H), 3.04 (m, 2H), 2.90 (s, 3H).

(S)-6-amino-2-(4-(((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)(methyl)carbamoyl)-1H-1,2,3-triazol-1-yl)hexanoicacid (GTJC-057)

To a solution ofN-((3R,4S,5R,6R)-4-(4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl)-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N-methylpropiolamide(100 mg 0.2564 mmol) in Toluene (5 mL), was added DIPEA (66.15 mg,0.5128 mmol), CuI (48.50 mg, 0.2564 mmol) and(S)-3-azido-7-((tert-butoxycarbonyl)amino)-2-oxoheptanoic acid (92.7 mg,0.3076 mmol) at 0° C. The reaction mixture was stirred at roomtemperature. After 12 h, the reaction mixture was quenched with in HCl(5 mL) then stirred for 30 minutes. Organic compound from the separatedaqueous layer was extracted with 5% MeOH in DCM (3×25 mL). The combinedorganic layers were washed with brine and dried over anhydrous Na2SO4.The solvent was removed under reduced pressure at 45° C. temperature toafford the title compound as a white solid (GTJC-057, single β isomer,15 mg, 5.2%).

HRMS (ESI) [M+H]+ calc. for C24H31 FN807 562.23, found: 563.38 [M+H]+

LCMS: m/z 563.3 (M+H)+ (ES+) 98.41% at 3.13 min.

1H NMR (400 MHz; DMSO-d6, single β isomer): □ 8.77 (s, 1H), 8.42 (m,1H), 7.68-7.75 (m, 2H), 7.38-7.57 (m, 1H), 7.12-7.16 (t, J=8.42 Hz, 1H),5.72 (d, 1H, J1-2=7.8 Hz, α-H-1), 5.64 (m, 1H), 5.25-5.35 (m, 1H),4.61-4.70 (m, 1H), 4.51 (bs, 1H), 3.93 (i, 1H), 3.71 (m, 1H), 3.54 (m,2H), 3.31-3.35 (m, 2H), 3.26-3.31 (in, 2H), 3.05 (s, 2H), 2.78 (in, 1H),2.49-2.54 (m, 2H), 2.44 (s, 1H), 2.13-2.24 (m, 1H), 1.55-1.59 (m, 1H),1.29 (m, 1H).

TABLE 1 Exemplary compounds with galectin-3 inhibition according to someembodiments Structures Inventory Code

G603 intermediate 1 Iminogalactose derivative

G606 intermediate 2 Carbogalactose derivative

G609 intermediate 3 Aminogalactose derivative

G611

G610

G617

G620

G622

G628

G629

G630

G631 GTJC-026

G632

G633

G635

G639

G642

G646

G647

G648

G649

G651

GS-052

G653

G655

G656

G657

G658

G662

G666

G667

G668

G669

G678

G679

G680

G637

1. A compound of formula 1 or a pharmaceutically acceptable salt orsolvate thereof:

wherein A is independently selected from the group consisting of NRa,CRb, PRc, and amino acid, wherein M is independently selected from thegroup consisting of NRa, CRb, PRc, ORd, SRe amino acid, and hydrophobichydrocarbons derivatives including heterocyclic substitutions of 3 ormore atoms, wherein Ra is selected from the group consisting of H, H2,CH3, COOH, NH2, COMe, halogen and combinations thereof, wherein Rb isselected from the group consisting of H, H2, O, OH, CH3, COOH, NH2,COMe, halogen and combinations thereof, wherein Rc is selected from thegroup consisting of O2, PO2, OH, halogen and combinations thereof,wherein Rd is selected from the group consisting of H, CH3, andcombinations thereof, wherein Re is selected from the group consistingof OH, O2, S, halogen and combinations thereof, wherein B is OH, NH2,NHAc, or NH-alkyl, wherein the alkyl group comprises 1 to 18 Carbons,wherein W is selected from the group consisting of O, S, CH2, NH, andSe, wherein Y is selected from the group consisting of O, S, NH, CH2,Se, S, P, amino acid, and hydrophobic linear and cyclic hydrophobichydrocarbons derivatives including heterocyclic substitutions ofmolecular weight of about 50-200 D and combinations thereof, wherein R₁,R₂, and R₃ are independently selected from the group consisting of H,O2, CO, NH2, SO2, SO, PO2, PO, CH3, linear hydrocarbon, and cyclichydrocarbon, and wherein the hydrocarbon is one of a) an alkyl group ofat least 3 carbons, an alkenyl group of at least 3 carbons, an alkylgroup of at least 3 carbons substituted with a carboxy group, an alkenylgroup of at least 3 carbons substituted with a carboxy group, an alkylgroup of at least 3 carbons substituted with an amino group, an alkenylgroup of at least 3 carbons substituted with an amino group, an alkylgroup of at least 3 carbons substituted with both an amino and a carboxygroup, an alkenyl group of at least 3 carbons substituted with both anamino and a carboxy group, and an alkyl group substituted with one ormore halogens, b) a phenyl group substituted with at least one carboxygroup, a phenyl group substituted with at least one halogen, a phenylgroup substituted with at least one alkoxy group, a phenyl groupsubstituted with at least one nitro group, a phenyl group substitutedwith at least one sulfo group, a phenyl group substituted with at leastone amino group, a phenyl group substituted with at least one alkylaminogroup, a phenyl group substituted with at least one dialkylamino group,a phenyl group substituted with at least one hydroxy group, a phenylgroup substituted with at least one carbonyl group and a phenyl groupsubstituted with at least one substituted carbonyl group, c) a naphthylgroup, a naphthyl group substituted with at least one carboxy group, anaphthyl group substituted with at least one halogen, a naphthyl groupsubstituted with at least one alkoxy group, a naphthyl group substitutedwith at least one nitro group, a naphthyl group substituted with atleast one sulfo group, a naphthyl group substituted With at least oneamino group, a naphthyl group substituted with at least one alkylaminogroup, a naphthyl group substituted with at least one dialkylaminogroup, a naphthyl group substituted with at least one hydroxy group, anaphthyl group substituted with at least one carbonyl group and anaphthyl group substituted with at least one substituted carbonyl group,d) a heteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group, and e) asaccharide, a substituted saccharide, D-galactose, Deoxygalactose,substituted D-Galctose, C3-[1,2,3]-triaZol-1-yl-substituted D-galactose,hydrogen, an alkyl group, an alkenyl group, an aryl group, a heteroarylgroup, and a heterocycle and derivatives, an amino group, a substitutedamino group, an imino group, or a substituted imino group.
 2. Thecompound of claim 1, wherein A-M is a spacer of at least 2 atomscomprising an amide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, amethylether —C(—H2)—O— methylester —C(═O)—O—, carbosulfon—C(—H2)-S(═O)(═O)—, phosphate —O—P(═O)(—OH)—, diphosphate—O—P(═O)(—O)—O—P(═O)(—O)—, Hydrazide —N(—H)—N(—H)—, amino acid, orcombinations thereof.
 3. The compound of claim 1, wherein A-M is aspacer linked to the anomeric carbon and comprises of an amide—N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, a methylether —C(—H2)-O—methylester —C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—, phosphate—O—P(═O)(—OH)—, diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—, carbohydrazide—C(═O)—NH—NH—, sulfonohydrazide —S(═O)2-NH—NH—, and phosphonicdihydrazide —P(═O)(—NH—NH2)(NH—NH—) spacer.
 4. The compound of claim 1,wherein A-M is a spacer of 2 or more atoms linked by single or doublebond: C—C, C═C, C—P, C—N, C—O, N—C, N—N, N═N, N—S, N—P, S—N, P—O, O—P,or combination thereof.
 5. The compound of claim 1, wherein the A-Mspacer comprises PO2 or PO2-PO2 bond linked to the anomeric carbon andto one or more atoms such as C or N or O or S.
 6. The compound of claim5, wherein C or N is linked to the anomeric carbon and PO2 or PO2-PO2 islinked to C or N.
 7. The compound of claim 1, wherein the A-M linked toR1 and R2 is N′-methylamide-3,4-difluorobenzene and Y—R₁ istriazole-3-fluorobenzene:


8. The compound of claim 1, wherein the A-M spacer is linked to agalactose, a hydroxyl cyclohexane, an aromatic moiety, an alkyl group,an aryl group, an amine group, or amide group.
 9. The compound of claim1, wherein the A-M spacer symmetrically links two galactosides orsubstituted derivatives thereof or wherein the A-M spacer asymmetricallylinks two galactosides or substituted derivatives thereof.
 10. Acompound or a pharmaceutically acceptable salt or solvate thereof having

wherein A-M is representing a spacer of at least 2 atoms comprising anamide —N(—Ra)—C(═O)—, sulfonamide —N(—H)—S(═O2)-, a methylether—C(—H2)-O-methylester —C(═O)—O—, carbosulfon —C(—H2)-S(═O)(═O)—,phosphate —O—P(═O)(—OH)—, diphosphate —O—P(═O)(—O)—O—P(═O)(—O)—,carbohydrazide —C(═O)—NH—NH—, sulfonohydrazide —S(═O)2-NH—NH—, andphosphonic dihydrazide —P(═O)(—NH—NH2)(NH—NH—) or combinations thereof,wherein A is independently selected from the group consisting of NRa,CRb, PRc, and amino acid, wherein M is independently selected from thegroup consisting of NRa, CRb, PRc, ORd, SRe amino acid, and hydrophobichydrocarbons derivatives including heterocyclic substitutions of 3 ormore atoms. wherein Ra is selected from the group consisting of H, H2,CH3, COOH, NH2, COMe, halogen and combinations thereof, wherein Rb isselected from the group consisting of H, H2, O, OH, CH3, COOH, NH2,COMe, halogen and combinations thereof, wherein Rc is selected from thegroup consisting of O2, PO2, OH, halogen and combinations thereof,wherein Rd is selected from the group consisting of H, CH3, andcombinations thereof, wherein Re is selected from the group consistingof OH, O2, S, halogen and combinations thereof, wherein B is OH, NH2,NHAc, or NH-alkyl wherein the alky comprises 1 to 18 Carbons, wherein Wis selected from the group consisting of O, S, CH2, NH, or Se, wherein Xis selected from the group consisting of O, N, S, CH2, NH, and PO2,wherein Y and Z are independently selected from the group consisting ofO, S, C, NH, CH2, Se, S, P, amino acid, and hydrophobic linear andcyclic hydrophobic hydrocarbons derivatives including heterocyclicsubstitutions of molecular weight of about 50-200 D and combinationsthereof, wherein R1, R2, R3, are independently selected from the groupconsisting of CO, O2, SO2, SO, PO2, PO, CH, Hydrogen, hydrophobic linearhydrocarbon, and hydrophobic cyclic hydrocarbon, wherein the hydrocarbonis one of: a) an alkyl group of at least 3 carbons, an alkenyl group ofat least 3 carbons, an alkyl group of at least 3 carbons substitutedwith a carboxy group, an alkenyl group of at least 3 carbons substitutedwith a carboxy group, an alkyl group of at least 3 carbons substitutedwith an amino group, an alkenyl group of at least 3 carbons substitutedWith an amino group, an alkyl group of at least 3 carbons substitutedwith both an amino and a carboxy group, an alkenyl group of at least 3carbons substituted with both an amino and a carboxy group, and an alkylgroup substituted with one or more halogens, b) a phenyl groupsubstituted with at least one car boxy group, a phenyl group substitutedWith at least one halogen, a phenyl group substituted with at least onealkoxy group, a phenyl group substituted with at least one nitro group,a phenyl group substituted with at least one sulfo group, a phenyl groupsubstituted with at least one amino group, a phenyl group substitutedwith at least one alkylamino group, a phenyl group substituted with atleast one dialkylamino group, a phenyl group substituted with at leastone hydroxy group, a phenyl group substituted with at least one carbonylgroup and a phenyl group substituted with at least one substitutedcarbonyl group, c) a naphthyl group, a naphthyl group substituted withat least one carboxy group, a naphthyl group substituted with at leastone halogen, a naphthyl group substituted with at least one alkoxygroup, a naphthyl group substituted with at least one nitro group, anaphthyl group substituted with at least one sulfo group, a naphthylgroup substituted With at least one amino group, a naphthyl groupsubstituted with at least one alkylamino group, a naphthyl groupsubstituted with at least one dialkylamino group, a naphthyl groupsubstituted with at least one hydroxy group, a naphthyl groupsubstituted with at least one carbonyl group and a naphthyl groupsubstituted with at least one substituted carbonyl group; and d) aheteroaryl group, a heteroaryl group substituted with at least onecarboxy group, a heteroaryl group substituted with at least one halogen,a heteroaryl group substituted with at least one alkoxy group, aheteroaryl group substituted with at least one nitro group, a heteroarylgroup substituted with at least one sulfo group, a heteroaryl groupsubstituted with at least one amino group, a heteroaryl groupsubstituted with at least one alkylamino group, a heteroaryl groupsubstituted with at least one dialkylamino group, a heteroaryl groupsubstituted with at least one hydroxy group, a heteroaryl groupsubstituted with at least one carbonyl group and a heteroaryl groupsubstituted with at least one substituted carbonyl group, e) saccharide,a substituted saccharide, D-galactose, substituted D-galactose,C3-[1,2,3]-triaZol-1-yl-substituted D-galactose, hydrogen, an alkylgroup, an alkenyl group, an aryl group, a heteroaryl group, and aheterocycle and derivatives; an amino group, a substituted amino group,an imino group, or a substituted imino group.
 11. A compositioncomprising a therapeutically effective amount of the compound of claim 1and a pharmaceutically acceptable adjuvant, excipient, formulationcarrier or combinations thereof.
 12. A composition comprising atherapeutically effective amount of the compound of claim 1 and atherapeutically effective amount of an anti-inflammatory drug, vitamin,pharmaceutical drug, nutraceutical drug, supplement, or combinationsthereof.
 13. A method of treatment of a disease in a subject in needthereof, comprising administering a therapeutically effective amount ofat least one compound according to claim
 1. 14. The method of claim 13,wherein the disease is a disorder related to pathological disease due toelevated galectin-3.
 15. The method of claim 13, wherein the disease isnonalcoholic steatohepatitis, fibrosis, cirrhosis, inflammatorydisorder, autoimmune disorder, neoplastic condition, metabolic disorderor cancer.
 16. The method of claim 15, wherein the inflammatory disorderis inflammatory bowel disease, Crohn's disease, multiple sclerosis,Systemic Lupus Erythematosus, arthritis, rheumatoid arthritis, asthma orulcerative colitis.
 17. The method of claim 15, wherein the fibrosis isliver fibrosis, kidney fibrosis, lung fibrosis, or heart fibrosis. 18.The method of claim 15, wherein the autoimmune disorder is rheumatoidarthritis or multiple sclerosis.
 19. The method of claim 13, wherein thedisease is heart failure, arrhythmias, uremic cardiomyopathy, chronickidney and lung diseases, skin autoimmune, or proliferative and fibroticskin disorder.
 20. The method of claim 15, wherein the neoplasticcondition is a benign or malignant neoplastic disease.